C2株贾第虫SBDS基因的原核表达与生物信息学分析  

Prokaryotic Expression and Bioinformatics Analysis of Giardia lamblia C2 Strain SBDS Gene

作　　者：王沂[1] 余源[2] 田喜凤[2] 李冀[2] 周英斌 李少东[2] 刘晓莉[2] 王洋[2] 

机构地区：[1]河北联合大学附属医院检验科,河北唐山063000 [2]河北联合大学生命科学学院,河北唐山063000

出　　处：《生物技术》2014年第5期28-33,共6页Biotechnology

基　　金：河北省青年科学基金项目("蓝氏贾第鞭毛虫α-4贾第素细胞定位和功能的研究";C2012401039)资助

摘　　要：目的:克隆、原核表达C2株蓝氏贾第鞭毛虫(Giardia lamblia,简称贾第虫)的Shwachman-Bodian-Diamond syndrome(SBDS),并对其进行生物信息学分析。方法:以C2株贾第虫基因组DNA为模板获得SBDS基因编码区,克隆入原核表达载体p ET-28a(+),测序并进行生物信息学分析,将重组质粒p ET-28a(+)-SBDS在大肠杆菌Rosetta(DE3)中诱导表达,SDSPAGE及Western blot验证表达效果。结果:成功构建了C2株贾第虫SBDS原核表达载体并在在大肠杆菌中得到了高效表达,SDS-PAGE及Western blot显示,在相对分子量约29k Da的位置出现目的蛋白条带,与理论值相符;生物信息学分析显示贾第虫SBDS蛋白在空间上形成三个结构域,在进化上与其它现存真核生物亲缘关系较远。结论:原核表达并分析了贾第虫SBDS蛋白,为贾第虫SBDS的进一步研究提供了基础。The Shwachman – Bodian – Diamond syndrome（ SBDS）,a highly conserved protein in Archaea and eukaryotes,is known to influence many cellular processes including ribosome biogenesis,mitotic spindle assembly,chemotaxis. In order to express Giardia lambia（ C2 strain） SBDS protein in E. coli,the full- length open reading frame of SBDS was amplified by PCR from Giardia lamblia genome DNA. The PCR product about 770 bp in length was cloned into prokaryotic expression vector p ET- 28a（ ＋）. Bioinformatics analysis showed that SBDS protein displayed three domains in three- dimensional space. Phylogenetic analysis showed a distant relationship of SBDS protein between Giardia lamblia and other eukaryotes. The recombinant SBDS protein was expressed in E. coli Rosetta（ DE3） by IPTG induction. SDS- PAGE and Western blot showed that the expressed product of SBDS was a fusion protein about 29 k Da. The successful prokaryotic expression and bioinformatics analysis of Giardia lamblia SBDS protein provide basis for further study of SBDS.

关 键 词：蓝氏贾第鞭毛虫 SBDS 原核表达 生物信息学 

分 类 号：Q786[生物学—分子生物学]