Cloning,Complementary Expression Vector Construction and Transformation of GbCBF2 Gene from Cotton

doi:10.7668/hbnxb.2016.01.007

Abstract

Abstract: In order to study the function of cotton CBF2 gene in plant resistance,the complementary expression vector pCAMBIA1300-AtCBF2P-GbCBF2-AtCBF2T was constructed,and transformed to Arabidopsis cbf2 mutant by genetic engineering.The four CBF2 genes were cloned from cDNA of the cotton Xinhai 16 and Arabidopsis CBF2 gene promoter AtCBF2P and termination AtCBF2T were also cloned by PCR method.Firstly,these genes were connected with pBluescript SK,and obtained the intermediate transition vector pBluescript SK-AtCBF2P-GbCBF2-AtCBF2T,then the AtCBF2P-GbCBF2-AtCBF2T fragment was obtained by cutting the intermediate transition vector with Sac Ⅰ and BamH Ⅰ,after the fragment inserted into pCAMBIA1300,the complementary expression vector pCAMBIA1300-AtCBF2P-GbCBF2-AtCBF2T was constructed.The cotton CBF2 gene was transformated into Arabidopsis cbf2 mutant by Agrobacterium mediated method and regenerative seedlings were obtained.These results will lay the foundation for screening cotton negative regulator CBF2 gene and stress-resistance molecular breeding.

Key words: Cotton, CBF2 gene, Complementary expression vector, Transformation

CLC Number:

Q78
S562.03

KONG Liying, LI Xiang, LI Caiyun, LIU Xiaodong, LI Yue. Cloning,Complementary Expression Vector Construction and Transformation of GbCBF2 Gene from Cotton[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2016, 31(1): 40-45. doi: 10.7668/hbnxb.2016.01.007.

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