Vitis amuerensis CBF3 Gene Isolation,Sequence Analysis and Expression  

作　　者：WANG Zhan-bin FENG Lian-rong WANG Jin-jie WANG Zhi-ying 

机构地区：[1]Northeast Forestry University/Key Labotatory of Forest Tree Genetic Improvement and Biotechnology, Ministry of Agricuture, Harbin 150040, P.R.China [2]Institute of Poplar of Liaoning Province, Gaizhou 115200, P.R. China

出　　处：《Agricultural Sciences in China》2010年第8期1127-1132,共6页中国农业科学（英文版）

基　　金：supported by the Fundamental Research Funds for the Central Universities,China(DL09EAQ02);the Natural Science Foundation of Heilongjiang Province and Harbin City,China(C200606nd and 2006RFQN005)

摘　　要：The full length cDNA sequence of CBF3 （CRT/DRE-binding factor） was cloned from Vitis amurensis by reverse transcription polymerase chain reaction （RT-PCR） using the primers designed based on CBF genes available in GenBank. Sequence analysis showed that the gene had 854 bp long and its coding sequence contained 720 bp, encoding a protein with 239 amino acids and an AP2 structural domain. The molecular mass of CBF3 was predicted to be 25.9 kDa and its theoretical isoelectric point was 7.02 and aliphatic index was 59.29. The average hydropathicity of the protein was -0.551. The tertiary structures of CBF3 were also analyzed. The prokaryotic expression vector pGEX-4T-CBF3 containing CBF3 gene was constructed and CBF3 fusion protein （52 kDa） was produced in Escherichia coli after induction with 1 mmol L-1 IPTG. Further studies are needed to evaluate its potential application for improving plant resistance to cold and other stress condition such as drought and salinity.The full length cDNA sequence of CBF3 （CRT/DRE-binding factor） was cloned from Vitis amurensis by reverse transcription polymerase chain reaction （RT-PCR） using the primers designed based on CBF genes available in GenBank. Sequence analysis showed that the gene had 854 bp long and its coding sequence contained 720 bp, encoding a protein with 239 amino acids and an AP2 structural domain. The molecular mass of CBF3 was predicted to be 25.9 kDa and its theoretical isoelectric point was 7.02 and aliphatic index was 59.29. The average hydropathicity of the protein was -0.551. The tertiary structures of CBF3 were also analyzed. The prokaryotic expression vector pGEX-4T-CBF3 containing CBF3 gene was constructed and CBF3 fusion protein （52 kDa） was produced in Escherichia coli after induction with 1 mmol L-1 IPTG. Further studies are needed to evaluate its potential application for improving plant resistance to cold and other stress condition such as drought and salinity.

关 键 词：Vitis amurensis CBF gene cloning sequence analysis expression in prokaryote 

分 类 号：Q785[生物学—分子生物学] Q78