农业生物技术学报

International Debate on Biosafety of Genetically Modified Crops:
Scientific Review on Several Cases of Debate

Jia Shirong Jin Wujun

2003, 11(1): 1-5 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Abstract: There were several cases of international debates on genetically modified crops: (1) Dr. Pusztai in 1998 claimed that genetically modified potato with snowdrop gna gene inserted had an adverse effect on the health of rat after feeding test. In the following year the Royal Society of UK conducted a thorough pure review on this issue. The conclusion indicated that the Pusztai's results were incorrect in many aspects, such as the experiment design, the execution and the statistics analysis etc. And no such an adverse effect should be drawn from it. (2) In 1999, 《Nature》 published a paper by one group from Cornell University in which they reported that the larvae of monarch butterfly were killed by the pollen of Bt corn on milkweeds. Subsequent laboratory and field studies have concluded that the larvae are not likely to be harmed by pollen of Bt corn in natural conditions. The fact is that the majority of pollens move only a short distance away from cornfield and cornfield typically contains a lower density of weeds, including milkweeds. The decline of population density of monarch butterfly is mainly caused by the over use of pesticides and the environmental changes occurred in Mexico. (3) In 1998, volunteer canola resistant to three herbicides was documented in a producer's field in Northern Alberta, Canada, which was then so called "super weeds" by activists. In fact, "super weeds" is not a scientific term and there is no such a thing existed in the nature as volunteer resistant to three herbicides can be killed by spray of herbicide 2,4-D. (4) In November 2001, Quist and Chapela published a paper on 《Nature》 claimed that DNA sequences similar to CaMV35S promoter and adh1 gene used in Bt 11 were found in the samples of maize landraces collected from Oaxaca, Mexico. It was then so called "gene pollution or gene contamination" by Green Peace. However, subsequent scientific analysis demonstrated that the sequence of 35S promoter was artifacts and the sequence of adh1 was the adh1-F that was a native gene in maize but not the transgenic gene of adh1-S used in Bt 11. CIMMYT has tested samples recently collected in Mexico fields and in their gene bank. The results showed that none of them contained 35S promoter sequence. (5) In June 2002, the Green Peace published a report saying that "Bt cotton damaged environment in China", which was totally flawed and misinterpreted. The positive consequences of Bt cotton grown in China have not been cited in Green Peace's report. The fact is, as a result of application of Bt cotton, the amount of pesticides used for control of cotton bollworm has been dramatically reduced by 70~80 percent. The population size of predators and the diversity of arthropods in Bt cotton field have increased. Because of less pesticides were used in the early stage of cotton growing season, the natural enemies of cotton aphids were significantly increased that resulted in a dramatic reduction of aphid population by 443~1546 folds in Bt cotton field comparing with that of in non-Bt cotton field. Monitoring bollworm population nationwide in cotton growing area has shownthat none of them have developed resistance to Bt powder or Bt cotton to date. The migration behavior of bollworm, the inheritance of insect resistance to Bt controlled by an incomplete recessive gene, the existing of "natural refugees" by multiple cropping systems in Northern China and the application of transgenic cotton with double genes (Bt/CpTI ), all of these have the functions in delay the development of resistance in bollworm population to Bt cotton. In conclusion, basically the nature of international debate on biosafety of genetically modified crops is not a pure scientific issue rather it is related to the economic and trade considerations.

Cloning and Prokaryotic Expression of GDA2 Gene and Purification of
GDA2-GST Fusion Protein

Sultan Ababakri1,2 Bai Yong1 Zhu Yuxian1**

2003, 11(1): 6-10 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Abstract: GDA2 is one of the senescence-related genes that were cloned from G2 pea (Pisum sativum L.) previously. It shares 36% sequence identity with the previously reported GDA1 gene. This cDNA was amplified by PCR method from the original cloning vector, pBSK-GDA2, and then cloned into the T-easy-vector to produce the new construction, pGEM-T-GDA2. Sequence analysis showed that the cloned cDNA of GDA2 is 1 120 bp long, which has an open reading frame of 642 bp and encodes a protein of 213 amino acids with a calculated molecular weight of 24 kD. By PCR method, Bgl Ⅱand XhoⅠ restriction sites were added to the 5' and the 3' end of the GDA2 respectively. The expression vector pGEX-GDA2 was constructed by inserting the coding region of GDA2 cDNA into pGEX 4T-1. And the IPTG-inducible product, a fusion protein which consists of 27 kD GST protein and 24 kD GDA2 protein at C-terminal, was finally purified by Glutathione Sepharose 4B affinity column.

Cloning and Characterization of a Stetaria italica Lipid Transfer Protein cDNA

Feng Xiaoyan Yu Jingjuan Zhao Qian Ao Guangming**

2003, 11(1): 11-15 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Abstract: A cDNA encoding lipid transfer protein (Siltp ) was isolated from Stetaria italica cDNA library. According to the results of Blastn and Blastp in GenBank, it shares 72%,70%,60%,54% nucleotide sequence identity and 79%,73%，65%,57% amino acid sequence identity with Hordeum vulgare lipid transfer protein gene LTP7a2b and LTPcw-19, Zea mays ltp and Brassica napus ltp separately. It was sequenced and characterized. Siltp contains an open reading frame of 363 bp, 121 amino acid. The first 26 amino acid is supposed to be a signal sequence. The mature protein has a molecular mass of 9.3 kD. Southern blotting analysis indicated that Siltp is one copy in the genome. Siltp expresses in stem, leaves and grains. Using software PAUP to analyze evolution of LTP from 17 different plants, phylogenetic tree allows the conclusion that LTP has four groups. Differentiation happened earlier in wheat，while in the other monocots such as maize， millet，barley and rice happened later. It shows the early gene replicate matters. Dicotyls scatter in phylogenetic tree which illuminates the evolution of LTP gene is complex. It shows LTP gene of corns evolved and divided from one ancestral LTP gene. Siltp has most identity with barley LTP in the phylogenetic tree.

Introducing Glucose Oxidase Gene into Rice Mediated by
Agrobacterium tumefaciens

Peng Hao1 Wang Zhixing2 Dou Daolong1 Zhu Shengwei 1 Lu Tiegang1 Sun Jingsan1*

2003, 11(1): 16-19 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Abstract: The glucose oxidase (GO ) gene with the function of broad-spectrum pathogen defense was inserted into a binary vector pCAMBIA1301 containing selectable marker of hygromycin resistant gene to produce a new expression vector, pCAG1301. The vector was introduced into Agrobacterium tumefaciens strain LBA4404 and then transformed them to immature embryos of rice (Oryza sativa L.ssp. japonica ) cultivar Nipponbare. Putative transformed plants were regenerated from hygromycin resistant calli, and the integration of GO gene was confirmed by Southern blotting analysis on rice genome. The H2O2 generated by the transgenic plants was detected by color reaction with starch-KI and this proved that the glucose oxidase generated by the expression of GO gene had exerted its function in rice. The transgenic plants were proved to have strong resistance to rice blast disease(Magnaporthe grisea ).

cDNA-AFLP Analysis of the Gene Expression
in Cotton Ovule at Fiber Initiation Stage

Xiao Yuehua Luo Ming Wei Yutuo Hou Lei Pei Yan**

2003, 11(1): 20-24 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Abstract: Using cDNA-AFLP technique, gene expressions in the ovules of cotton (Gossypium hirsutum L.) Xuzhou142 (WT) and its lint and fuzz and mutant (fl ) at fiber initiation stage(0 and 4 d post anthesis) were compared. The result showed that among 561 amplified bands, 166 belonged to two kinds of ovules, of which 83 specially displayed in WT ovules and 83 in fl , respectively. Using cDNAs as probe, 47 WT special fragments were further analyzed by reverse Northern blot, of which 32 were found to be differentially expressed in the WT ovule, and presumably related to the fiber initiation. Sequence analysis demonstrated that, among 18 sequenced fragments, eight were sequence-similar to proteins in GenBank, i.e. polyphosphoinositide binding protein, cytochrome oxidase subunit Ⅱ, beta-glucosidase, sucrose synthase, calcium channel alpha-1D subunit, rice osmotic response protein osr40 and TSO1-like protein, four were similar to hypothetical proteins of Arabidopsis thiliana , and no similar sequence were found in GenBank for the other 6 fragments.

Molecular Cloning and Expression of EIN3 (Ethylene Insensitive 3)
Gene in Tomato Fruit

Xu Jingyu Luo Yunbo** Wei Shaochong

2003, 11(1): 25-29 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Abstract: EIN3(ethylene insensitive 3) in Arabidopsis thaliana is a novel nuclear-localized protein, which is thought to serve as a transcription factor in response to ethylene signal. The cDNA of homologous genes of A.thaliana EIN3 was obtained from tomato (Lycopersicon esculentum ) fruit by RT-PCR amplification and cDNA library screening，termed Le-EIL . The cDNA clone contained an open reading frame of 1 845 bp , encoding a sequence of 615 amino acid. Primary sequence analysis indicated that it exhibited 79% sequence similarity to tobacco(Nicotiana tabacum ?雪 TEIL and the sequence similarity of this clone to A.thaliana EIN3, EIL1,EIL2 and EIL3 was 59%、56%、52%、and 46% respectively. Its expression patterns in both wild type tomato and antisense-ACS transgenic tomato fruit development showed that the expression of Le-EIL gene highly relatived to ripening and ethylene synthesis. In wild type tomato fruit, the expression of Le-EIL gene increased as the fruits got ripening, and reached its peak level at turning and pink stage, then decreased. In antisense-ACS transgenic tomato fruit, the expression of Le-EIL was weak at 60 d fruit past anthesis but disappeared at the other ripening stages.

Molecular Marker Location of a Novel Yellow Rust Resistance Gene
YrSp Derived from Spelt Wheat

Wei Yanling Ni Zhongfu Xie Chaojie Yang Tsomin Sun Qixin** Du Jinkun

2003, 11(1): 30-33 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Abstract: The losing of varietal resistance to yellow rust(Puccinia striiformis Westend f. sp. tritric ) is an important factor causing periodical and massive yellow rust outbreak in wheat growing area. Emergency and development of new isolates of yellow rust pathogen have put widely planted varieties in China into a serious crisis of resistance losing and therefore put searching for new resistance resources into great need. Molecular marker technology could facilitate the identification and location of novel genes. The current study was undertaken using SSR technology in order to identify a novel yellow rust resistance gene derived from Spelt wheat (Triticum spelta L.) and study its chromosomal location. After screening 150 wheat microsatellite primers, a marker Xgwm155-147 bp located on chromosome 3A linked to this gene with the genetic distance of 40.5 cM was found, which indicated that the resistance gene is located on chromosome 3A. Since the only yellow rust resistance gene Yr5 known to be derived from Spelt is located on chromosome 2BL, the resistance gene identified in this study might be a novel yellow rust resistance gene which is temporarily designated YrSp.

Expression, Chromosomal Location and Molecular Marker of High-Molecular-Weight Glutenin Subunit Gene of
Thinopyrum elongatum in Wheat Background

Tang Zhaohui Liu Shoubin You Mingshan Li Baoyun Mao Shanfeng Song Jianmin Liu Guangtian

2003, 11(1): 34-39 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Abstract：High-molecular-weight glutenin subunits (HMW-GS) are main factors that affect bread-making quality of common wheat. Many common wheat cultivars derived from the Thinopyrum germplasm have good bread-making quality. But HMW-GS coded by E-genome of Thinopyrum was not reported previously. In the paper, the HMW-GS compositions of Thinopyrum elongatum (2n=14, EE) were analyzed by SDS-PAGE technique with materials of wheat (Triticum aestivum) var. Chinese Spring (CS), Thinopyrum elongatum (PI547326), CS-Th. elongatum amphidiploids, alien disomic and ditelosomic addition (DA) and alien disomic substitution (DS) lines. Studies on chromosomal location of HMW-GS gene of E-genome was performed in the meantime, and works of amplifying the HMW-GS gene of E-genome was also conducted by using PCR primers specific for the repetitive region of y-type HMW-GS genes. The main results were showed bellow:
1. E-genome in CS background coded one single HMW-GS with a highly similar migration ratio to subunit 1By8 of CS in SDS-PAGE gel, and in the accession of Thinopyrum elongatum PI547326 coded one subunit similar to 1By9 of Chenny. These two subunits were therefore temporarily named 1E8 and 1E9 respectively. 2. The HMW-GS gene of E-genome was located in the long arm of chromosome 1E. This result was consistent with the fact that the arrangement of HMW-GS genes was collinear in chromosomes of grain crops. 3. With primers for HMW-GS genes, one specific fragment in the approximately length of 1 300 bp was amplified in all materials containing 1EL chromosome. This fragment could be used as molecular marker for the HMW-GS gene of E-genome.

bFGF Gene Expression in Goat Ovary Follicular Cells and
Partial Sequence Analyses of bFGF cDNA

Wang Baoli1,2 Zhang Yong1

2003, 11(1): 44-48 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Abstract: A RT-PCR method for detecting gene expression of the goat basic fibroblast growth factor (bFGF) was set up. The expressions of bFGF gene in goat ovarian follicle, cumulus-oocyte complexes (Caprane) (COCs), granulosa cells(GCs) and theca follicular cells were observed, respectively. The results indicated that all these cells expressed bFGF mRNA. Meanwhile, the bFGF cDNA was amplified. The isolated fragment (283 nt) was cloned and sequenced. Comparison of the nucleotide sequence indicated 99.3% similarity to the bovine bFGF gene and 95.4% similarity to the human bFGF gene.

Regeneration of Interspecific Somatic Hybrid between
Sweetpotato and Its Wild Relative Ipomoea triloba*

Wang Jingshan1 Liu Qingchang2** Meng Xiangxia1 Wang Weihua1 Xu Lijuan1 Zhai Hong2

2003, 11(1): 40-43 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Abstract: Protoplasts of sweetpotato (Ipomoea batatas ) cv. Genki were fused with protoplasts of its wild relative I. triloba by the PEG method. The fusion products were cultured in a modified liquid MS medium containing 0.05～0.20 mg/L 2,4-D and 0.5 mg/L KT. After 7 weeks of culture, small calluses 1～2 mm in size were transferred to solid MS medium supplemented with 0.05～0.20 mg/L 2,4-D and 0～0.5 mg/L KT for callus proliferation. When the calluses were transferred either to the basal medium or to the medium supplemented with 3.0 mg/L BAP and further to the basal medium, 69 plants were regenerated. PCR analysis indicated that 1 of 69 regenerated plants was the somatic hybrid. This somatic hybrid showed the climbing habit looked like I. triloba and the stem color, green to purple, looked like Genki. The internode length and the leaf shape were intermediate. The chromosome number of root tip cells ranged from 54 to 103, depending on different root tip cells.

Expression of Bacillus circullans Lactase Gene in E. coli
and Properties of Expressed Lactase

Wang Yuanhuo1 Yao Bin1** Yuan Tiezheng1 Cao Shishu1 Zhang Wei2 Wang Yaru1 Fan Yunliu2

2003, 11(1): 83-88 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Abstract : The lactase gene from Bacillus circulans was overexpressed in Escherichia coli and the expressed lactase had normal bioactivity. The properties of purified lactase expressed in E. coli and produced by Bacillus circulans were determined. The results revealed, between the lactase expressed in E. coli and produced by Bacillus circulans, that there had notable difference in enzyme properties in optimal pH value and pH stability, optimal temperature and temperature stability, Km value. The optimal pH value of the lactase expressed in E. coli was 5.0, whereas that of origin lactase produced by Bacillus circulans was 6.5. The optimal temperature of expressed lactase was 37 ℃ and 55 ℃, whereas that of origin lactase was 55 ℃. After treated for 2 h under pH 4.0, the remaining enzyme activity of expressed lactase was 85%, whereas that of origin lactase was only 5%. The thermal stability of expressed lactase was better than that of origin lactase, after treated for 10 minutes under 60 ℃, the remaining enzyme activity of expressed lactase was 30% , whereas origin lactase almost lost all of its activity. The expressed lactase had better metal ion resistance to Cu2+, Mn2+ and Fe2+, the remaining enzyme activity of expressed lactase and origin lactase in reaction system with 1mmol/L Fe2+ was 96% and 22%, respectively. The Km and Vmax values of expressed lactase had 285-fold decrease and 5.4-fold increase, respectively, compared to those of origin lactase. The lactase was overexpressed in E. coli, the expression level of lactase was high up to 33.102 U/mL. The properties of the lactase expressed in E. coli including high activity in pH 3.5～8.5, good thermal stability and metal ion resistance demonstrate that the enzyme is a good candidate in dairy industry.

Cloning of cDNA Fragment of an Antennal-specific Gene in Helicoverpa armigera

Wang Guirong Guo Yuyuan Wu Kongming**

2003, 11(1): 49-54 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Abstract: The olfactory sensors of insects' antenna, which directly exposed to the environment, play an important role in identifying volatile hydrophobic molecules in the air. However, many odors in the air are cytotoxic xenobiotics, so a certain mechanism must exist in insect's antenna to decompose cytotoxic xenobiotics and redundant odorant stimulators. In the present study, differential display polymerase chain reaction (dd-PCR) was used to isolate three antennal -specific mRNA from the moth of cotton bollworm(Helicoverpa armigera ). Sequence analysis indicated that A4-1 clone was homologous to the glutathione S-transferase family of proteins. Northern blot showed that A4-1 clone was specifically expressed in the antenna of H. armigera and the expressed level was higher in male antenna than that in female ones, which was similar to the expression of PBP-Harm gene from cotton bollworm. According to the gene's expression in insects' antenna and the role of glutathione S-transferase reported in other insects, the gene was predicted to be responsible for the decomposition of pheromone and harmful xenobiotics in antenna.

Comparative of Three Methods for the Induction of Tetraploidy Pearl Oyster (Pinctada martensi D.)

Wang Aimin 1,2 Yan Bing 2 Lan Guobao2 Ye Li 2

2003, 11(1): 64-69 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Abstract: Three methods for the induction of tetraploidy pearl oyster (Pinctada martensi D.) were compared. 1. Inhibiting release of the first polar body in fertilization of eggs from triploidy with sperm from diploidy resulted in the formation of tetraploid larvae at 27.7% ~ 35.9 %. But the death rate was high and no living tetraploid was detected after the larval stage. 2. Tetraploidy was induced by inhibition of the first and the second polar bodies in normal diploid zygotes. There were over 30% tetraploid in D-staged larva and the death rate was high. But at mid- oyster stage, i.e. 7 months cultivation, about 0.0625 % were tetraploid. 3. Inhibition of the first cleavage of normal diploid zygotes resulted in approximately 13.1% tetraploids at trochophore stage . However, the hatching rate was low and the death rate was high. Tetraploidy were not found after trochophore stage . The studies indicated that Inhibiting formation of the first and the second polar bodies in normal diploid zygotes could produce living tetraploid pearl oysters .

Improvement of a Method for Determining and Differentiating Virulence of Infectious Bursal Disease Virus (IBDV) Isolates and the Virulence Test of 7 Chinese IBDV Isolates

Zhu Yiping1 Zhang Manfu1** Chen Guanchun2 Wang Xiaomei2 Wang Xiurong2 Thierry van den Berg 3
Dolores Morales3 Boonleong Lim4

2003, 11(1): 55-59 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Abstract: A precise evaluation of virulence of infectious bursal disease virus (IBDV) is very important in studying the regulation of virulence variation and therefore is also important in finding the solutions to IBDV infection. However, the virulence determination method is not standardized. Normally, the virulence of IBDV is expressed as the mortality rate of SPF chicken or commercial chicken with variable inoculation dose, and the virus is titrated accordingly with TCID50、ELD50 or EID50 et al. The aim of the study was to unify IBDV titration route and standardize the method of IBDV virulence determination by greatly increasing the sensitivity of EID50 with chorioallantoic membrane (CAM) inoculation and high sensitive double-sandwich ELISA, hence made it possible to titrate all pathotypes of IBDV by EID50. The inoculation dose was 2×103 EID50 according to the survey report by Eterradossi. The standardized method was proved to be high sensitive, good reproducibility and universal adaptable by the virulence test of 7 Chinese isolates.

Construction and Identification on cDNA Library of Breast
Muscle and Liver in Silkie Fowl

Wang Xiuli1,3 Li Ning2** Zhao Xingbo1 Zhao Zhihui2 Deng Xuemei1
Li Changlü１ Qiu Xuemei3 Wu Changxin1

2003, 11(1): 60-63 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Abstract: More and more animal breeding scientists are interested in chicken economic trait research. Most of these traits are related to different gene expression. So understanding the expressed genes and constructing the known gene expressed profile from muscle tissue and liver tissue of chicken are very important in animal breeding and genetics research. In order to prepare for the future comparative genomics research, functional genomics research, QTLs mapping, and animal molecular breeding etc, we have successfully constructed a breast muscle tissue cDNA libraries and a liver tissue cDNA libraries of Silkie Fowl (Gallus gallus ) in the study. The vector of cDNA used was Lambda ZAP II. At the same time, both cDNA libraries' quality were identified by the PCR for phage, the enzyme of EcoRⅠand XhoⅠ digested for plasmid and other methods. The results showed that the OD260/OD280 of mRNA from breast muscle and liver were 1.92 and 1.90 respectively, the titer of the breast library was 3.6×107 pfu/mL，the recombinant percentage was 92%，and the average length of inserted cDNA fragments was longer than1.5 kb. The titer of the liver cDNA library was 3.4×107 pfu/mL，the recombinant percentage was 90%，and the average length of inserted fragment cDNA was longer than 1.4 kb.

Genetic Linkage Map Construction of Some Microsatellites on Pig
Chromosome 4 and 7

Zuo Bo Xiong Yuanzhu Su Yuhong Deng Changyan Yu Li Zheng Rong

2003, 11(1): 70-74 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Abstract: A reference family was established by using three Large White boars (Sus scrofa )and seven Meishan dams(S.scrofa ). Blood samples of 140 F2 individuals were collected. Each sample was amplified by using primers of 7 microsatellites loci on chromosome 4 and 8 microsatellites loci on chromosome 7, and genotypes were identified. The linkage map of chromsome 4 and 7 was constructed by using CRIMAP software. In this population, average allele number was 3.6, average heterozygosity and polymorphic information content 0.531 and 0.463, average information meiosis 235. The genetic length of sex-average, female and male maps were 178.5, 197.7 and 172.2 cM for chromosome 4, and 179.1, 195.7 and 167.7 cM for chromosome 7 respectively. The average marker interval of chromosome 4 and 7 was 29.9 and 24.9 cM, respectively. Construction of linkage map will make the foundation for QTL mapping afterwards.

Construction and Expression of the Specific Secretory Vector
in the Chicken Oviduct Cells

Sun Mingjun Zhao Chen Sha Jin Han Haitang Zheng Kejian Li Zandong**

2003, 11(1): 75-78 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Abstract:To express foreign proteins in chicken (Gallus gallus ) oviduct cells and secret to oviduct, 450 bp chicken lysozyme cDNA fragment was cloned by using RT-PCR method. Treated by the restriction endonuclease Bam HⅠand Hind Ⅲ, the fragment was ligated with the vector generated by the same Bam HⅠ-HindⅢ digestion of plasmid pOVA-GFP. The resulting expressive vector pOVALG included an 1.2 kb chicken ovalbumin 5'-flanking regulatory region and a lysozyme-GFP fusion gene with the signal peptide of the lysozyme. After transfecting the vector to the chicken embryo fibroblasts and mammary cancer cell line of rat SHZ-88 cells, green fluorescent protein had been detected successfully.

Cloning and Expression of a Baculovirus J Domain Protein
Gene in E. coli and Preparation of Antiserum

Wang Lihua Yu Jianxiu Yin Chong Li Zhaofei Pang Yi**

2003, 11(1): 79-82 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Abstract: Spodoptera litura multicapsid nucleopolyhedrovirus bjdp (baculovirus J domain protein ) gene is the only gene encoding a J domain protein homology in known baculovirus genome. Sequence analysis indicated that the N terminus of the gene production had a conserved J domain. With the specific primers which designed on bjdp gene reported recently, the coding region was amplified from SpltMNPV genome DNA via PCR . The PCR product was cloned into the pGEM-T Easy vector to get the recombinant plasmid (pGT-B). The bjdp gene was recombined in vitro with expression vector pQE30 and transformed into E.coli M15[pREP4]. The M15[pREP4] containing bjdp recombinant plasmid expressed a 36 kD 6×His-tag fusion protein after the induction with 1mmol/L IPTG. The fusion protein was purified with Ni-NTA resin column and used as the immunogen to raise a BJDP-specific antiserum. Western blot analysis indicated that the antiserum could react with the corresponding protein existed in SpltMNPV-infected S1-zsu-1 cells.

Molecular Identification of the Mutant Isolates of Magnaporthe grisea

He Yueqiu1,2 Hei LEUNG2 Robert S. ZEIGLER2 Tang Wenhua3

2003, 11(1): 89-93 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Abstract: Pathogenic mutant isolates 9 from the greenhouse and 5 from the International Rice Research Institute Blast Nursery, respectively, were identified by using molecular techniques with Rep-PCR of the two primer pairs, Pot2-1/Pot2-2 and PMC1-2/PMC1-3, RAPD of 177 primers and RFLP of MGR586. The results indicated that the 9 mutants from greenhouse had almost identical DNA band patterns and their original isolates lost 1~3 bands for 7 mutants, but no changes for the other 2 mutants. The 5 mutants from the nursery, which defeated resistance genes Pi-1 and Pi-2, originated from the isolates with avirulence gene Avr-1 in the field, not from outside by genetic shift based on their pathotypes and DNA band patterns. Therefore, it was concluded that the mutation was a main evolution force for Magnaporthe grisea.

Detection of the Polymorphisms of Growth Hormone Gene in
Chinese and European Pig Breeds

Wang Wenjun1，2 Chen Kefei1 Ren Jun1 Gao Jun1 Ding Nengshui1 Li Lin1 Huang Lusheng1**

2003, 11(1): 103-104 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Obtain To Transgenic Color Cotton of Expressing Insect-Resistant Gene

Wang Dongmei1 Li Jianping1 Huang Leping1 Li Renjing2**
Guo Jiangyong2 Wu Minggang2 Tian Yingchuang3 Guo Hongnian3

2003, 11(1): 101-102 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Preparation and General Properties of Streptomyces flavovirens A8
Immobilized on Silk Fibroin

Zhu Xiangrui Xu Junliang

2003, 11(1): 105-106 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Preliminary Study on the Plasmids of Thermus Strains from Hot Springs in
Guantang Qionghai Hainan

Wang Ruiping Ran Ling Wu Haitao Xing Xuezhong

2003, 11(1): 107-108 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Difference of Nuclear and Cytoplasmic in Vitro Maturation of
Mammalian Oocytes and Related Important Factors

Li Yonghai Liu Ruihua Jiao Lihong Wang Hong Wang Weihua**

2003, 11(1): 94-100 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Abstract: Techniques of oocyte maturation in vitro affect the developmental competence of oocyte fertilization in vitro and embryo culture in vitro directly. It was reported that the requested environment or conditions were different between nuclear and cytoplasmic maturation in vitro. The criterion of nuclear maturation was indicated by the extrusion of first polar body and the oocyte reaching to metaphase Ⅱstage but there was no accepted standard for evaluating cytoplasmic maturation so far. Most of the researchers worldwide took the developmental competence of oocyte as their cytoplasmic maturation evaluating system. In general, cytoplasmic maturation of oocyte needs better conditions and longer culturing time than that of nuclear maturation in vitro. The conditions and results of oocyte maturation in vivo and in vitro, animals treated with hormones and some other factors affecting oocyte maturationin vitro and development in recent reports were discussed in the paper.