农业生物技术学报

Relationship of Lignin Content with the Stem Color in the
Transgenic Poplar with Depressed Expression of 4CL Gene

2004, 12(6): 621-624 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Abstract:The lignin content of transgenic triploid Chinese white poplar (Populus tomentosa Carr.) expressing the antisense 4CL (4-coumarate:CoA ligase) cDNA was remarkably reduced. The stems of transgenic poplars were complete red-brown or mottled red-brown immediately after the bark was peeled off, while the stems of the untransformed plants were white. The reductions in lignin content of transgenic plants with complete red-brown stems were much higher than with mottled red-brown ones. Wiesner reactions of untransformed plants were typical purple-red, while that of complete red-brown stems of transgenic plants exhibited red. Wiesner reactions of white parts in mottled red stems of transgenic plants displayed typical purple red like untransformed plants, while that of red-brown parts showed red color, which testified further the relationship of lignin content with stem color in transgenic plant. It was considered that the stem color of transgenic plant could be employed as supplementary trait to screen the transgenic plant with the reduced lignin content.

β-glucosidase cDNA Cloning in the Tea(Camellia sinensis )
and Its Prokaryotic Expression

　LI Yuan-Hua JIANG Chang-Jun** YANG Shun-Li YU You-Ben

2004, 12(6): 625-629 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Abstract：The β-glucosidase gene has important effects on the alcoholic aroma precursors and insect-resistance. The complete cDNA squence of β-glucosidase of tea(Camellia sinensis) was cloned and its full length was 1 475 bp (GenBank, Accession No. AF537127), and shared 40%～60% similarity to corresponding parts of β-glucosidase gene from other plants in nucleotide sequence. Its secondary structure contained 14.33% α-helical conformation, 25.43% β-sheet conformation and many function domains of amino acid. The β-glucosidase gene was cloned into the pET-32a expression vector and expressed high-efficiently in Escherichia coli BL21(DE3), and the molecular weight of expressed fusion protein was 63 kD. The results of emzymatic reaction showed that fusion protein possesed normal bioactivity, and it could catalyze the dehydration of the glycodic bond. The fusion protein was mainly expressed by soluble protein in cytoplasm.

　　Structural Analyses of Functional Region of Maize
In5-2 Promoter in Transgenic Tobacco

　　YUAN Yuan LIANG Ben-Guo LIU Yun-Jun WANG Tao**

2004, 12(6): 630-634 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Abstract: The different length of six maize In5-2 promoter 5' end deletions were obtained by PCR method. The mutants were cloned to plant expression vector D1301 and transferred into Nicotiana tabacum. GUS activity analysis showed that regulation region of its basic transcription was mainly within -1 520 to -1 431 bp upstream of ATG. DNAMAN analysis there were two stem-loop structures within -1 108 to -1 079 and –891 to -864 bp related to ATG. It was suggested that their protein integration site was between -1 079 to -897 bp related to ATG. In addition, cis-acting elements responsive to chloroacetanilide could be mainly located within the range of-388 to -86 bp upstream of ATG.

S1 Nuclease Sensitivity of Surrounded Nucleolus(SN)- and Non Surrounded Nucleolus(NSN)-type Mouse Ooctyes during In vitro Maturation

LI Jun YANG Ming-Sheng LI Ying-Xiang XU Yin-Xue LIU Hong-Lin**

2004, 12(6): 658-661 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Abstract: The fully grown mouse germinal vesicle oocytes from large antral follicle can be classified into two groups, according to their chromatin configuration. The S1 nuclease sensitivity of them was investigated during their maturation in vitro. The results showed that NSN-type (non-surrounded nucleolus, whose decondensed chromatin distributed in almost whole nuclear) oocytes at MⅠ stage had more S1 nuclease sensitivity than that of SN-type (surrounded nucleolus，whose condensed chromatin surrounded nucleolus.) oocytes (P <0.01), while little difference between them in S1 nuclease sensitivity was found at MⅡ stage (P >0.05). It seemed that the NSN-type oocytes at MⅠ stage had more abundant single-strand property than SN-type oocytes, and during developing from MⅠ stage to MⅡ stage, the abundance of single-strand character in these two-type oocytes became homogenized.

Expression of Recombinant Chicken IFN-γ Gene in Baculovirus Vector System

WANG Hai-Yan2 LIU Sheng-Wang1** KONG Xian-Gang1 ZHAO Zhen-Hua2

2004, 12(6): 639-643 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Abstract：The cDNA of ChIFN-γ gene was subcloned into BamHⅠ site of pMelBacB under the control of baculovirus polyhedrin to construct recombinant transfer vector pMelBacB-ChIFN-γ. The recombinant transfer vector pMelBacB-ChIFN-γ was then transfected into sf9 insect cells with wild-type Bac-N-Blue linear DNA using cationic liposomes. The cultures were harvested and plaque-assayed with x-gal, and blue plaques were selected as recombinant viruses. After being purified for three times and identified by PCR, the purified viruses were named rBaculovirus-ChIFN-γ. The Sf9 cells were infected with rBaculovirus-ChIFN-γ and incubated at 28 ℃, their and cultures were harvested at 24, 48, 72, 96, 120 and 144 h post-inoculation, respectively. The lysates of the cells were analyzed by SDS-PAGE, a band with a apparent molecular weight of 19 kD was observed. To further confirm this band, the lysates of sf9 insect cells infected with rBaculovirus-ChIFN-γ were analyzed by Western blot using polyclonal antibodies against ChIFN-γ, the result showed that this band represented ChIFN-γ. It was demonstrated that about 19 kD recombinant ChIFN-γ protein with activity was expressed in sf9 insect cells.

Development and Preliminary Application of the gE Enzyme Linked Immunosorbent Assay for Detecting Antibodies to gE Protein of Pseudorabies Virus in Pigs

TANG Yong1,2　　 CHEN Huan-Chun1** 　　QIN Ya-Li1 　　HE Qi-Gai1 　　JIN Mei-Li1　　 WU Bin1 　　LIU Zheng-Fei1

2004, 12(6): 635-638 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Abstract: To differentiate pigs infected with pseudorabies virus (PrV) and vaccinated with Ge-PrV, the gE enzyme linked immunosorbent assay(ELISA) based on the recombinant glycoprotein E, which was expressed by E .coli, purified, denatured and renatured, was developed. The diagnostic specificity and sensitivity of the developed gE-ELISA were evaluated by testing 115 serum samples. The gE-ELISA's diagnostic specificity was 94.5% and sensitivity was 96.7%. Five serum samples were tested with plates from 5 patches and a coefficient of variation of less than 10% was obtained, showing the gE-ELISA had good reproducibility. When gE-ELISA was compared with a commercial blocking ELISA by testing 356 serum samples, the agreement rate of the two assays was 92.13%(328/356). The above results indicated that the gE-ELISA developed in our laboratory could be used to differentiate the PrV infected and gE-PrV vaccinated pigs.

Develeopment of Bovine Somatic Cell Nuclear Transfer and
Parthenogenesis Embryos in Two-step Culture System In vitro

LI Yu-Qiang ZHANG Xiu LI Yu ZHANG Yong

2004, 12(6): 653-657 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Abstract: The development of bovine parthenogenesis and somatic cell nuclear transfer embryos in different culture systems was studied. In vitro maturated bovine oocytes were activated by 5 mmol/L ionomycin for 5 min treatment and 2 mmol/L 6-DMAP for 4 h culture. The nuclear transplantation embryos were produced by intracytoplasmic injection of the bovine skin fibroblast nuclear. The results indicated that the blastocyte rate was higher when bovine parthenogenesis embryos cultured in BECMaa (bovine embryo culture media) + 10% FBS than in SOFaa + 10% FBS or TCM199 + 10% FBS (16.5%∶13.6% / 12.8%, P＜0.05); However, adding 1 mmol/L glutathione into SOFaa + 10% FBS could significantly increase parthenogenesis blastocyte rate (18.5%∶12.8%, P＜0.01). When added into serum-free SOFaa (including GSH) the fibroblast growth factor 4 (FGF4) and insulin could improve bovine nuclear transfer embryo development significantly respectively (8.7% / 9.3%:5.5%, P ＜0.05), but there was no obviously effect when FGF4 was used together with FBS. According to the above results and metabolism character of early bovine embryo, the two-step bovine embryo culture system in vitro, which embryos were cultured firstly in media Ⅰ(SOFnaa + EDTA + GSH + insulin) for 72 h, then transferred into media Ⅱ (SOFaa + 5％FBS + Glucose + GSH + insulin) for 8 d, was established. As compared with one-step culture using media SOFaa + 10％FBS + GSH, two-step culture system could significantly improve bovine parthenogenesis and nuclear transfer embryos development, whichever the blastocyte rate (22.3%∶18.5%; 15.0%∶11.7%, P＜0.01) or the development rate from 8/16-cell stage to blastocyte(47.3%∶43.6%; 37.4%∶32.5%, P＜0.05). In conclusion, the two-step culture system was a feasible bovine embryos culture method in vitro, and much more accorded with the development character and nutrition demand of bovine early embryo.

Rat Adipocyte Apoptosis Induced by Tumor Necrosis Factor-α

LIN Ya-Qiu LI Rui-Wen SUN Chao CHEN Guo-Zhu
YANG Yong-Qing MENG De-Lian YANG Gong-She**

2004, 12(6): 644-647 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Abstract: The effects of tumor necrosis factor-α(TNF-α) at different concentration levels on rat adipocyte apoptosis were detected by optical microscopy, agarose gel electrophoresis and flow cytometry methods. The morphologic changes of rat adipocyte apoptosis induced by TNF-α were correlated linearly with the concentrations of TNF-αwithin a range from 5 to 20 ng/mL, and the higher concentration of TNF-α could induce more obvious apoptosis. The morphologic changes of rat adipocytes treated by 5 ng/mL TNF-α were clearly shown, but their DNA ladders didn't appear by the DNA electrophoresis analysis, indicating that the morphologic changes occurred earlier than the biochemical changes. TNF-α induced the rat adipocyte apoptosis in a dose-dependent manner within a concentration range from 5 ng/mL to 20 ng/mL. Compared with the control, the apoptotic effect induced respectively by 5, 10, 15 and 20 ng/mL TNF-α was significantly difference (P<0.01), but the effect among 10, 15 and 20 ng/mL TNF-α treatments was not difference (P >0.05). Thus the optimum concentration of TNF-α inducing apoptosis was determined at the level of 10 ng/mL.

Separation and In vitro Culture of the Goat Fetal Testicular Cells
and the Behavior of Male Germ-line Stem Cells(mGSCs)

DONG Wu-Zi1 HUA Jin-Lian1 ZHUANG Shu-Zhen1 SHEN Wen-Zheng1,2 DOU Zhong-Ying1**

2004, 12(6): 648-652 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Abstract： The seminiferous tubules tissues from goat fetus were treated with 4 methods. The treatment with 0.1% colleagenase for 15 min and then with 0.25% trypsin for 5 min showed ideal cell dispersal degree after centrifugation and washing. The in vitro culture of the cells separated with different medium system showed that mulberry-shaped mGSCs clusters and a single layer of Sertoli cells appeared as original generation in 120 h, the mGSCs clusters developed half-suspendedly and distributed in different locations with the Sertoli cells in the plate; The cells in the margin area of incompletely digested mGSCs clusters and the single cells from them were obviously homogenized after culture for 5 days in fiber cell feeding layer from mice fetus, while same-treated mGSCs clusters co-cultured with Sertoli cells did not displayed clear homogenization in marginal cells of the clusters in the same culture system; And the first generation of mGSCs clusters co-cultured with Sertoli cells formed the same morphologic properties with the original generation of clusters, cells in clusters were tight and mGSCs divided slower than one in MEF feeding layer.

Polymorphism of Jiangxi Indigenous Chicken Breeds and Its
Population Genetic Relationships Inferred by AFLP Analysis

HU Xiao-Fen1，3** GAO Jun1** AI Hua-Shui1 DING Neng-Shui1 CHEN Cong-Ying1
GUO Yuan-Mei1 SHU Xi-Fan2 HUANG Lu-Sheng1 REN Jun1***

2004, 12(6): 662-667 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Abstract: A total of 15 pairs of AFLP primer combinations were used to detect genetic variation of pooled genomic DNA from 8 native chicken breeds and 1 cultivated chicken breed in Jiangxi Province as well as 1 exotic chicken breed, which included Nanchenghei, Yuganwuhei, Congrenma, Ningdusanhuang, Wanzaikanglehuang, Guangfeng Baierhuang, Dongxiang green-shell, Taihe silky fowl, Jingdehuang and Israel recessive white feather fowl. The genetic similarity coefficients of 10 chicken breeds were calculated from AFLP data, the highest similarity coefficient of 0.7790 existed between Yuganwuhei and Nanchenghei, followed by that of 0.6726 between Guangfeng Baierhuang and Jingdehuang, while the lowest similarity coefficient of 0.2801 was revealed between Israel recessive white feather fowl and Congrenma. UPGMA (unweighted pair group method with arithmetic mean) cluster analysis was also performed by the program of PHYLIP. Eight Jiangxi indigenous chicken breeds and the cultivated breed (Jingdehuang) were grouped into one branch, in which Yuganwuhei and Nanchenghei clustered together, and Guangfeng Baierhuang and Jingdehuang were grouped closely, while exotic chicken breed (Israel recessive white feather fowl) formed another branch. The results indicated that Jiangxi indigenous chicken breeds and exotic chicken breed had distant genetic relationship, the closest geneticrelationship was observed between Nanchenghei and Yuganwuhei, which was consistent with the previous result by RAPD analysis. The two chicken breeds were therefore suggested to be combined for conservation. Guangfeng Baierhuang and Jingdehuang showed intimate relationship, the two breeds was closer than any other pair of breeds, which was also consistent with the RAPD result and the breeding history of Jingdehuang chicken breed using Guangfeng Baierhuang as one of founder animals.

Developmental Changes of GnRH-I and ER-β mRNA
Expression in the Ovary of Prepubertal Shaoxing Ducks

NI Ying-Dong ZHOU Yu-Chuan LU Li-Zhi CHEN Jie ZHAO Ru-Qian**

2004, 12(6): 668-671 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Abstract: Gonadotrophin-releasing hormone-1(GnRH-I )and estrogen receptor-β (ER-β ) mRNA expression in the ovary of Shaoxing ducks between 1 and 90 post hatching were measured by semi-quantitative RT-PCR using β-actin as an internal standard. The abundance of ovarian GnRH-I mRNA increased gradually from 1 to 90 d of age, 60-day-old ducks exhibited the highest level of ovarian GnRH-I mRNA. And the level of ovarian GnRH-I mRNA from 60 to 90 day was significantly higher than that of 1 to 30 d. However, the level of ER-β mRNA went up gradually from 1 d to 60 d, reaching a peak at 60 d, then went down to the level of 90 d. Ovarian expression of ER-β mRNA was significantly greater at 60 d than any of other age groups. The results indicated that ovarian GnRH-1 may be involved in regulating the growth of post-stage ovarian follicle, while ER-β mRNA may be participated in the earlier follicle development in Shaoxing ducks.

Optimization on Process of Transgenic Mice Production

ZHENG Min2 WANG Li-Li1 WANG Mei-Li2 NIU Hui-Ling2 DAI Yun-Ping1*

2004, 12(6): 672-675 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Abstract: The microinjection and implantation conditions were optimized by Kunmin mice as donor and receipt in the production of transgenic mice. Simultaneously the DNA concentrations in different size and standard of receipt selecting were redressed. Eventually the positive rates of transgenic mice were improved from 10% to 20%; the optimum suitability of DNA concentration was 2~3 μg/mL when size of DNA fragment exceed 10 kb.

Cloning and Expression of Spodoptera litura nucleopolyhedrovirus
gp41 Gene in Escherichia coli and Preparation of Its Antibody

PAN Li-Jing LI Zhao-Fei YIN Chong LV Lei PANG Yi **

2004, 12(6): 676-679 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

　Abstract: GP41, a major glycoprotein, has been identified in the Occlusion-derived virus（ODV）of baculoviruses, which was required for the egress of nucleocapsids from the nucleus in the pathway of the Budded virus（BV）synthesis. The ORF of Spodoptera litura nucleopolyhedro virus（SpltMNPV）gp41 gene was obtained by PCR method from SpltMNPV genomic DNA. The PCR product was cloned into pMD18-T vector to get the recombinant plasmid（pT-gp41）. The gp41 gene was recombined in vitro with prokaryotic expression vector pQE30 and transformed into Escherichia.coli M15 [pREP4]. The M15 [pREP4] strain, containing gp41 recombinant plasmid, expressed a 37.9 kD 6×His-tag fusion protein after induction with 1 mmol/L IPTG (isopropylthio-β-D-galactoside). The fusion protein was purified with Ni-NTA resin column and used as the immunogen to raise a GP41-specific antibody. Western blotting analysis indicated that the antibody was suitable to be used for further analysis of GP41 protein.

Cloning and Sequencing of DNA Components of
Banana bunchy top virus Hainan Isolate

TIAN E ZHUANG Jun LIU Zhi-Xin*

2004, 12(6): 680-684 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Abstract：Six DNA components of Banana bunchy top virus （BBTV）were amplified by PCR using total DNA from banana tissues which are of characteristic of typical BBTV symptom as template. The sequence analysis indicated that the 6 fragments were all full-length components of BBTV, they were 1 104, 1 067, 1 059, 1 045, 1 014 and 1 081 nt respectively. All of them have been submitted to GenBank(Accession No. AY450396, AY606084, AY494786, AY494788, AY606085 and AY494787, respectively). The viral sequences from different countries were compared. Results revealed that the sequence of component 1 was more conservative than other components, DNA1 of Hainan isolate shared homologies of 95.4% with that of Vietnam isolate, while both the sequences of component 2 and component 4 had more difference, and DNA2 of Hainan isolate only shared homologies of 83% with that of Guangzhou isolate. According to DNA1 sequence comparison results, the BBTV Hainan isolate should belong to Asia sub-group.

Primary Analysis of Molecular Diversity in Populations of the
Fungus Ustilago scitaminea Syd.

QUE You-Xiong XU Li-Ping** ZOU Tian-Tang CHEN Ru-Kai

2004, 12(6): 685-689 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Abstract: Eighteen isolates were obtained from sugarcane smut fungus (Ustilago scitaminea Syd). cultivated in six different provinces or regions including Fujian Province. On the basis of isolation of single spore and the acquisition of series of the isolates, 13 random primers of 10 bp were occupied in analysis of genetic diversity by RAPD. The largest dissimilarity coefficient was 0.89474, being observed in two isolates with codes of 7 and 9 originated from Fujian and Guangxi, respectively; the smallest value of 0.25217 was observed in two isolates originated in Fujian. Dendrogram of UPGMA cluster analysis revealed that 18 isolates of U. scitaminea Syd. were divided into 6 groups according to the dissimilarity coefficient of 0.75. Because of small dissimilarity coefficient, seven isolates obtained from sugarcane in Fujian Province belonged to two adjacent groups of cluster Ⅰ and cluster Ⅱ. High dissimilarity coefficient of 0.82353 was observed in two isolates collected from same host origin but with different geographical origin.The results of cluster suggested that the molecular diversity was associated with geographyic origins in some degree, but not related to host origin.

Isolation and Identification of Nitrogen-fixing Bacilli

DING Yan-Qin1 WANG Jian-Ping1 LIU Yuan1 Chen San-Feng1** WANG You-Shan2 XING Li-Jun2

2004, 12(6): 690-697 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Abstract: A total of 29 isolates were selectively obtained from the rhizospheres of wheat(Triticum aestivum ), maize(Zea mays ), ryegrass(Lolium sp. ) and willow(Salix sp.) based on their growth on nitrogen-free medium and their resistance to 100 ℃ for 10 min. 7 out of them had nifH gene determined by PCR amplification and belonged to the genera Bacillus and Paenibacillus based on their physiological and chemical characteristics, 16S rDNA sequence (GenBank accession No. AY373358, AY373360~AY373364 and AY376876), G+C mol% and DNA-DNA hybridization. And the isolate T1 was identified as Bacillus cereus. The isolates G1, C4 and C5 were identified as Bacillus megaterium. The isolates W5 and G2 were simlar to Bacillus marisflavi and Paenibacillus polymyxa in physiological and chemical characteristics, 16S rDNA and G+C mol%, respectively. The isolate T7 might represent a novel species of Paenibacillus.

Construction of Antagonistical, Insecticidal and Endophytic Multifunctional-engineering Strains by Protoplast Fusion

YAN Nian-Long1 QIU Si-Xin2 HE Hong3 GUAN Xiong1 HU Fang-Ping2**

2004, 12(6): 704-708 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Abstract： A multifunctional-engineering strain was constructed by protoplast fusing of the insecticidal Bacillus thuringiensis with the endophytic B. subtilis. The results of PCR, insecticidal and endophytic tests showed that the vip3A gene was existed in the fusant, the antagonism of the fusant was against the Fusarium oxysporum f.sp.cucumerinum and the adjusted mortality to the Plutella xylostella reached 59%, at the same time, its endophytic character was also existed.

Effects of E26 Agent on the Expressions of
Defense Genes pal and sts of Grape Suspension Cells

ZHENG Zu-Yu LI Jin-Yun WANG Jian-Hui WANG Hui-Min**

2004, 12(6): 698-703 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Abstract: Using phenylalanine ammonia-lyase (PAL) and stilbene synthase (STS) as indicators of grape resistance to pathogen infection, the effects of different components (bacterial cells, fermentation products and metabolites) of strain E26(Agrobacterium vitis ), biocontrol agent of crown gall, on PAL activities and genes' transcriptions of pal and sts in suspension cells of two grape varieties Vitis romanetii cv. Qiuhe and V. vinifera cv. Cabernet Sauvignon) were studied. The PAL activities of Qiuhei and Cabernet Sauvignon were higher than the controls at 12 h to 48 h after treated by E26 bacterial cells, 4 h to 48 h after treated by E26 fermentation products and 12 h to 48 h after treated by E26 metabolites. Dot blotting analysis showed that the quantities of pal and sts transcriptional products (mRNA) both in Qiuhei and Cabernet Sauvignon were higher than that of the control at 12 h to 48 h after treated by E26 bacterial cells; At 4 h to 48 h after treated by E26 fermentation products, the quantities of pal transcriptional products in Qiuhei were higher than that of the control, while that in Cabernet Sauvignon were 4 h to 12 h. The quantities of sts transcriptional products in the two varieties were higher than that of control at 4 h to 48 h after treated by E26 fermentation products; At 4 h to 48 h after treated by E26 metabolites, the quantities of pal transcriptional products in Qiuhei were higher than that of the control, while that in Cabernet Sauvignon were 4 h to 24 h. The quantities of sts transcriptional products in the two varieties were higher than that of the control at 6 h to 48 h after treated by E26 metabolites.

Effect of Buffer Systems on Elastase Production by Bacillus sp. EL31410

XU Ying1 HE Guo-Qing1* CHEN Qi-He1 LI Jing-Jun2

2004, 12(6): 709-713 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Abstract：The elastase is a kind of protease that can hydrolyse elastin. pH is one of the most important environmental factors for cell growth and product accumulation. The best pH for cell growth and elastase production were about 6.0～7.0 and 6.4～6.6, respectively. Single factor and rotatable orthogonal central composite design were used to study the effect of buffer systems on producing elastase. The optimal result showed that 15 h later adding pH 6.4 (0.02 mol/L KH2PO4-NaOH) buffer to medium was the best way for cell growth and elastase production. This model was verified to be credible.

Expression and Purification of GST-GFMCry11B Fusion Protein in E.coli and Preparation of Polyclonal Antibody Against GFMCry11B

DING Min1,2 WANG Feng1** SU Jun1 CHEN Zhi-Hou1 CHEN Zai-Jie1 SUN Ming2

2004, 12(6): 714-719 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Abstract: The coding region of GFM cry11B, fully synthesized gene, was cloned into the multi-clone sites of pGEX4T vector to produce the recombined expression plasmid pGC by the methods of restriction enzyme digestion. The recombinant plasmid was transferred into Escherichia coli BL21(DE3). GST-GFMCry11B fusion protein was obtained after adding IPTG into the growth media. SDS-PAGE analysis revealed that the fusion protein was highly expressed and accumulated up to above 21% of the total bacterial protein and the main expression production was deposited as inclusion body. The GST-GFMCry11B was purified with Glutathione Sephrose 4B after soluble treatment. GFMCry11B protein cleaved by Thrombin was collected and used as the antigen to immune the rabbits. ELISA assay showed that the titer of the prepared polyclonal antibody was 1∶8000 and had high specialties.

Obtaining Transgenic Fertility Pear (Pyrus communis L.) Plants with
Antifungal γ-thionin Rs-afp1 Gene

ZHAO Rui-Hua1 LIU Qing-Zhong2 SUN Qing-Rong2 ZHANG Xian-Sheng1

2004, 12(6): 729-730 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Cloning and Sequence Analysis of α Subunit of Phycoerythrin Gene from Corallina officinalis

WANG Sheng ZHONG Fu-Di WU Zu-Jian LIN Qi-Ying XIE Lian-Hui**

2004, 12(6): 733-734 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Mapping of Sterile Gene of Pingxiang Dominant Genic Male Sterile Rice Using SSR Technique

HE Hao-Hua1 LIU Xiao-Qiang2 ZHU Chang-Lan1 PENG Xiao-Song1 WAN Jian-Min3 WANG Xiang-Kun4

2004, 12(6): 727-728 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Semisynthesis of Insulin-like Growth Factor-I for Transforming Cabbage
and Construction of MAR-regulated Expression Vector

ZHANG Ying-Hua1，2 WANG Xiao-Jia1**

2004, 12(6): 731-732 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

RAPD Analysis of Tobacco (Nicotiana tabacum L. cv. K346) Leaf Number and Trunk High Mutant

TANG Yong-Hong1 JIA Jing-Feng2 CHEN Gang2

2004, 12(6): 735-736 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Cloning and Characterization of a Protective Antigen
H11 cDNA of Haemonchus contortus

YAN Ruo-Feng LI Xiang-Rui*

2004, 12(6): 739-740 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Comparision of Tephritidae's Genomic DNA Extraction Methods

YU Jie1 CHEN Jin-Song 2 WENG Rui-Quan2 QIU Jun-Zhi1 GUAN Xiong 1*

2004, 12(6): 741-742 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

A Method for RNA Isolation from Pear Style

XU Yi-Liu ZHANG Shao-Ling**

2004, 12(6): 737-738 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Simultaneous Detection of Two Orchid Viruses by Dual One-step RT-PCR

ZHOU Guo-Hui1 CHEN Xiao-Qin1 LI Mei-Hui2 ZHOU Jie-Lang3 TANG Tian-Hua3 FENG Sheng-Xiang3
GUO Li-Jing3 , ZHANG Wang3

2004, 12(6): 743-744 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Research Advance on Molecular Mechanism of Odors Perception in Insects

WANG Gui-Rong WU Kong-Ming** GUO Yu-Yuan

2004, 12(6): 720-726 | doi: | Full text (HTML) (1 KB) | PDF

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Abstract

Abstract: The olfactory behavior of insects is very complicated and involves many kinds of proteins, namely, odorant-binding proteins, odorant degrading enzymes and odor receptors etc.. Some recent advances on them and chemo-electrical signal transduction are reviewed.