Study on the Transformation of Antisense Soybean Fat Acid Desaturase to Agrobacterium tumefacien

doi:10.7668/hbnxb.2011.02.005

Abstract

Abstract: In this study,the primers were designed based on the sequences of soybean fat acid desaturase(fad2-1)in NCBI database to amplify fad2-1 gene from the genomic DNA of soybean leaves.The amplicon was cloned into pMDl8-T vector to be introduced to E.coli JMl09.and then was sequenced and aligned with the sequence of soybean fad2-J in NCBI database,and reversely inserted in pBt expression vector to construct plant antisense expression vector,which was mobilized into Agrobacterium tumefacien strains LBA4404 by freeze-thawing method to get genetically modified strain LBA4404 confirmed by double enzyme digestion and PCR detection.The results indicated that the size of the isolated gene Was 1 196 bp,bearing 95% identity with that in NCBI database showing that it was soybean fad2-1,and that antisense fad2-1 expression vector was successfully constructed and transferred into Agrobacterium t.m,facien strains LBA4404.This will build up the foundation for the improvement in soybean faty acids by antisense teehnique.

Key words: Glycine max(L．)Merr, Fat acid desaturase gene, Antisense technique, Agrobacterium tumefacien, Transformation

CLC Number:

ZHOU Yan-qing, TIAN Miao-miao, WANG Fang, CHEN Yan-mei, ZHANG Yong-hua, YUAN Bao-jun. Study on the Transformation of Antisense Soybean Fat Acid Desaturase to Agrobacterium tumefacien[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2011, 26(2): 23-25. doi: 10.7668/hbnxb.2011.02.005.

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