SARS病毒S2蛋白在毕赤酵母中的表达及鉴定  

Expression of Severe Acute Respiratory Syndrome(SARS) Virus S2 Protein in Pichia pastoris

作　　者：王瑞琳[1] 金宁一[2] 赵博[3] 贾雷立[2] 金洪涛[2] 金明兰[2] 屈勇刚[2] 

机构地区：[1]吉林大学畜牧兽医学院,长春130062 [2]军事医学科学院十一所病毒室,长春130062 [3]吉林大学生命科学学院,长春130023

出　　处：《中国生物制品学杂志》2006年第6期560-563,共4页Chinese Journal of Biologicals

基　　金：吉林省科技厅重大项目(20030413).

摘　　要：目的研究用毕赤酵母菌表达SARS病毒S2蛋白亚单位。方法依据GenBank中SARS基因组序列,采用人工合成的方法合成SARS病毒刺突蛋白S2亚单位的基因(1590bp),再与CTL表位基因(195bp)重组后,将其克隆到毕赤酵母表达载体pPIC9K中,构建重组表达质粒pPIC9K-S2,转化毕赤酵母GS115,并经MD和MM平板筛选及PCR鉴定,得到阳性重组酵母工程菌GS115/pPIC9K-S2。经诱导、表达,并对表达产物进行分析、鉴定。结果所构建的重组质粒pPIC9K-S2正确,在毕赤酵母中可高效表达,其表达产物与阳性SARS血清产生特异性反应。结论SARS病毒刺突蛋白S2亚单位可在毕赤酵母中高效表达,产物具有良好的抗原特异性。Objective To express severe acute respiratory syndrome（SAILS） virus S2 protein in Pichia pastoris. Methods A 1 590 bp gene fragment encoding the S2 protein of SARS virus was synthesized according to the SARS virus genome sequence reported in GenBank and recombined with CTL epitope gene（ 195 bp） ,then cloned into Pichia pastoris expression vector pPIC9K. The constructed recombinant plasmid pPIC9K-S2 was transformed to Pichia pastoris GS115 ,and the positive clones were screened by MD and MM plate cultures and identified by PCR. The obtained recombinant Pichia pastoris strain GS115/pPIC9K-S2 was induced with methanol, and the expressed product was identified by SDS-PAGE and Western blot. Results Recombinant plasmid pPIC9K-S2 was correctly constructed and highly expressed in Pichia pastoris. The expressed product showed specific reaction with SARS virus-positive serum. Conclusion SARS virus S2 protein was successfully expressed in Pichia pastoris and showed good antigenic specificity.

关 键 词：重症急性呼吸系统综合征(SARS) S2蛋白 毕赤酵母 

分 类 号：R563.1[医药卫生—呼吸系统]