BnRCH同源基因耐热性分析  

The thermostability analysis of the BnRCH homologous genes

作　　者：唐天元[1] 陈存[1] 陈凤莲[1] 杨毅[1] 

机构地区：[1]四川大学生命科学学院生物资源与生态环境教育部重点实验室,成都610064

出　　处：《四川大学学报（自然科学版）》2012年第5期1169-1174,共6页Journal of Sichuan University(Natural Science Edition)

基　　金：国家自然科学基金(30971557);转基因专项(2009ZX08009-080B)

摘　　要：本实验室先前通过酵母双杂交在油菜中获得一个未知基因(BnRCH),NCBI保守区域查寻表明BnRCH蛋白合有一个RINGv(RING-variant)结构域,该结构参与多种抗逆途径.为初步探索BnRCH的作用,通过RT-PCR技术,克隆BnRCH及其在其它物种中的同源基因cDNA序列,之后将各基因克隆到原核表达载体pET-28a(+)中进行表达.通过IPTG诱导,western检测表明重组蛋白的相对分子量约为35 kD.进一步在高温环境(43℃)对其生长曲线测定,发现重组菌生长状态优于对照菌,说明BnRCH及其同源基因能提高微生物的耐热性.An unknown gene（BnRCH）in B. napus was found by yeast two hybrid system, and NCBI Conserved Domain Search reveals that the BnRCH protein contains a RING-variant domain （RING-variant） ,which is involved in many degeneration-resistance pathways. In order to explore the function of BnRCH, BnRCH gene and its homologous sequences from other species were amplified by RT-PCR, and then cloned into the prokaryotic vector pET-28a（＋）for expressing. After IPTG induction, Western blot analyses showed that the relative molecular mass of expressed recombinant proteins were about 35 kD. Furthermore, the growth curve test under high temperature（43 ℃） illustrates that the growth status of recombinant bacteria is better than that of the control bacteria, which shows that BnRCH and its homologous genes possess the feature of enhancing microbe＇s thermal resistance.

关 键 词：BnRCH 耐热 大肠杆菌 生长曲线 

分 类 号：Q93[生物学—微生物学] Q78