百日咳毒素S1亚单位缺失分子的构建及其在重组杆状病毒中的表达  

Expression of truncated S1 subunit of pertussis toxin in recombinant baculoviruses

作　　者：李庆雷[1] 于康震[2] 马丽英[3] 王孝铭[3] 

机构地区：[1]哈尔滨医科大学病生教研室,哈尔滨150001 [2]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室,哈尔滨150001 [3]哈尔滨医科大学病理生理教研室,哈尔滨150086

出　　处：《中国免疫学杂志》2000年第1期12-14,共3页Chinese Journal of Immunology

基　　金：国家自然科学基金!39580022

摘　　要：目的：为探明校基端疏水区对百日咳毒素Ｓ１亚单位分泌性的影响，试图构建缺失羧基端疏水区的Ｓ１亚单位。方法：应用ＰＣＲ技术构建了６种不同信号序列修饰的缺失羧基端疏水区的Ｓ１亚单位编码基因，并使用重组杆状病毒（ｒＢＶ）技术实现了这些缺失Ｓ１亚单位（ｔＳ１）在昆虫细胞中的高水平表达。结果：这些缺失Ｓ１亚单位的表达量为每万个昆虫细胞１．０１～２．２５ｕｇ。细菌信号肽和流感病毒ＨＡ信号肽均能完整表达和正确剪切，但ＨＡ信号肽在昆虫细胞中的处理效率更高。结论：ｔＳ１亚单位缺失羧基端疏水区后在昆虫细胞中的表达量明显提高，这些ｔＳ１亚单位的异源性表达是由于信号肽剪切不全所导致，ｔＳ１分子表观分子量的漂移并不意味着信号肽剪切不正确。Objective:In order to investigate the effect of the bydrophobic domains on secretion of the subunits,we deleted the bydrophobicdomains from the S1 subunits.Methods: Six signal modified S1 encoding DNA fragmants were constructed that lackal C-terminal hydrophobgicsequences using PCR technique expecting to get secreted S1 Proteins. Results: Those truncated S1 (tS1) subunits were abundantly expressed byrecombinant baculoviruses in insect cells (expression levels ranged from 1 .01 to 2.25 ug/104 cells) .The HA sighal could be processed more anciently than the native one, although they both could be cleaved correctly. Conclusion: The expression levels of the tS1 subunits are increased significantly after the deletion of the hydrophobic domains. The heterologous of the tS1 Proteins is the result of incomplete signal cleavage,and the apparent MW shifting does not imply incorrect signal cleavage.

关 键 词：百日咳毒素 S1亚单位 重组杆状病毒 信号肽 

分 类 号：R516.6[医药卫生—内科学]