摘要
目的 克隆和高效表达查菲埃立克体 (Ehrlichiachaffeensis) 2 8kD表面抗原基因。 方法 依据已知的查菲埃立克体 2 8kD表面抗原基因设计引物 ,用PCR从查菲埃立克体基因组中克隆该基因片段 ( 5 3 1bp) ,再将该基因片段定向插入表达载体 (PinPointTMXa - 3 )中 ,然后将该重组质粒转化E .coliJM10 9细胞。结果 经SDS -PAGE分析 ,转化子所产生的融合蛋白分子量为 3 2kD ,与预期的结果相一致 ;在 3 7℃用 0 .1MmIPTG诱导转化子 12h ,融合蛋白量达到细菌总蛋白量的 3 5 .8%。结论 查菲埃立克体 2 8～kD表面抗原基因在E .coliJM10 9细胞内得到高效率表达 ,从而为该抗原的大量制备奠定了基础。
Aim To clone and express the 28kD surface antigen gene of Ehrlichia chaffeensis.Methods With a pair of primers designed based on the known gene sequence of 28kD surface antigen of Ehrlichia chaffeensis,a gene fragment (531bp) was amplified from its genomic DNA by PCR.The fragment was inserted into PinPoint TM Xa-3 expression vector,and E.coli JM109 cells were transformed with the recombinant plasmid.Results By SDS-PAGE,the molecular weight of fusion protein produced by the transformant was detected to be 32kD,identical to that designed previously.The yield of fusion protein was 35.8% of the total proteins of the transformant under a 12h inducing with 0.1 mM IPTG at 37℃.Conclusion The 28kD surface antigen gene of E.chaffeensis is efficiently expressed in the E.coli cells,which lays a foundation of preparing a large amount of the antigen.
出处
《中国人兽共患病杂志》
CSCD
北大核心
2001年第6期14-17,共4页
Chinese Journal of Zoonoses
基金
国家自然科学基金资助项目 (批准号 :39870 0 42 )
