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| Construction of Plant Expression Vector pCAMBIA2301-del-ros and Genetic Transformation in Strawberry(Fragaria ananassa Duch.) |
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Abstract In order to study multi-gene interaction and reduce the blindness of many transformation, CaMV 35S promoter and nos terminator were added into gene del and ros upstream and downstream, respectively. del and ros genes can regulate anthocyanin synthesis. Intermediate vector pBI121-del and pCAMBIA1301-ros were constructed. BamHⅠand BglⅡproduced the same end tetra-nucleotide segments. The CaMV 35S promoter separately driven del and ros genes were constructed in plant expression vector pCAMBIA2301 under ligase. Strawberries(Fragaria ananassa Duch.) were treated by the constructed vector pCAMBIA2301-del-ros via Agrobacterium tumefaciens LBA4404 mediated transformation method. Transgenic strawberry lines were screened by PCR analysis. Observed by organizations, the roots and leaves of transgenic strawberry lines became reddish purple. The expression of anthocyanin biosynthetic genes anthocyanidinsynthase(ANS), chalcone-flavanone isomerase(CHI), flavanone 3-hydroxylase(F3H), dihydroflavonol-4-reductase(DFR) and UDP glucose-flavonoid 3-O-glcosyl-transferase(UFGT) were all enhanced in transgenic strawberry plant. The results were further confirmed that the plant expression vector pCAMBIA2301-del-ros could be used to transform plants. Construction of plant expression vector pCAMBIA2301-del-ros can provides a good idea for constructing a multi-gene plant expression vector and a method for studying interaction of multiple genes.
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Received: 01 November 2013
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HUANG Xing, YAN Ai-Fen, DENG Ting-Xian, OUYANG Hong-Jia, LIU Lian, FENG Juan, ZHU Xiang-Xing, NIE Qing-Hua, TANG Dong-Sheng, ZHANG Xi-Quan. Construction of Zinc Finger Nuclease-induced Targeting Vector of Luchuan Pig (Sus scrofa) Fat1 Gene and Transgenic Study In vitro[J]. 农业生物技术学报, 2019, 27(8): 1369-1381. |
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