核农学报2011，Vol.25Issue(2)：247-252,6.
印度芥菜BjJ10-2基因原核表达载体构建及抗盐功能的鉴定
CONSTRUCTION OF RECOMBINANT PROKARYOTIC EXPRESSION VECTOR FOR Brassica juncea BjJ10-2 AND IDENTIFICATION OF ITS FUNCTION FOR HIGH SALINITY TOLERANCE
摘要
Abstract
Using RT-PCR, we cloned the open reading frame (ORF) of RAMP4 super family member BjJ10-2 from Brassica juncea, then the prokaryotic expression vector pET32a-BjJ10-2 was constructed and transformed into E. coli strain BL21 (DE3)plysS. SDS-PAGE gel electrophoresis showed that BjJ10-2 was expressed in BL21 strain after IPTG induction. Using spectrophotometer we tested the salt tolerance ability of recombinant BL21 ( pET32a-BjJ10-2 ) and control BL21 (pET32a). The results showed that over-expression of BjJ10-2 markedly increased salt tolerance of BL21 compared with the control of empty vector transformed BL21 strain, and the critical concentration of NaCI resistance could be up to 0. 8mol/L. Semi-quantitative RT-PCR showed that BjJ10 - 2 was responded to salt stress, and its transcript abundance was significantly enhanced in the root of B. juncea and reached the peak at 8 ～ 12h after NaCl treatment. The result showed that BjJ10 -2 is a new salt stress responsive gene and might play an important role in the physiological processes of salt tolerance in plants.关键词
印度芥菜/NjJ10-2/原核表达载体/大肠杆菌BL21菌株/抗盐性/半定量RT-PCRKey words
Brassica juncea / BjJ10 - 2/ prokaryotic expression vector/ E. coli BL21/ salt resistant/ semi-quantitative RT-PCR引用本文复制引用
郝梦雨,郎明林,杨学举..印度芥菜BjJ10-2基因原核表达载体构建及抗盐功能的鉴定[J].核农学报,2011,25(2):247-252,6.基金项目
中国博士后科学基金(No.20060390479),河北农大博士启动基金(200805),转基因生物新品种培育重大科研专项基金(No.2009ZX08002-012B) （No.20060390479）
