| 伪狂犬病病毒闽A株糖蛋白gD基因去信号肽片段在大肠埃希氏菌中的表达 |
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| 引用本文： | 娄高明,邓汉虹,杨承槐.伪狂犬病病毒闽A株糖蛋白gD基因去信号肽片段在大肠埃希氏菌中的表达[J].中国兽医科技,2005,35(9):675-678. |
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| 作者姓名： | 娄高明 邓汉虹 杨承槐 |
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| 作者单位： | [1]韶关学院英东生物工程学院,广东韶关512005 [2]广东省汕尾市粮油食品进出口公司养猪场,广东汕尾516621 [3]中国兽医药品监察所,北京100081 |
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| 基金项目： | 国家“十五”科技攻关项目（2002BA514A-1-3）;广东省自然科学基金重点项目（020391） |
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| 摘 要： | 应用PCR方法从包含伪狂犬病病毒（PRV）Fa株糖蛋白gD基因的重组质粒pB13中扩增出糖蛋白gD基因去信号肽片段，将其克隆到大肠埃希氏菌表达载体pThiohisA中。测序结果表明，糖蛋白gD基因去信号肽片段长1155bp，与PRVFa株糖蛋白gD基因完全一致。经1mmol／L IPTG诱导后，重组质粒pThiohisA-gD在大肠埃希氏 菌XL1-blue、Top10、DE3和DH5a中均得到表达，其中在宿主菌XL 1-blue中表达量最高。Western-blotting结果显示，大肠埃希氏菌XL1-blue表达的糖蛋白gD具有免疫原性。
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| 关 键 词： | 伪狂犬病病毒 gD基因去信号肽片段 大肠埃希氏菌 诱导表达 免疫原性 |
| 文章编号： | 1000-6419（2005）09-0675-04 |
| 收稿时间： | 2005-08-08 |
Expression of gD gene encoding area of pseudorabies virus Fa strain exclude signal peptide in Escherichia coli |
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| Lou GaoMing;Deng HanHong;Yang ChengHuai.Expression of gD gene encoding area of pseudorabies virus Fa strain exclude signal peptide in Escherichia coli[J].Chinese Journal of Veterinary Science and Technology,2005,35(9):675-678. |
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| Authors: | Lou GaoMing;Deng HanHong;Yang ChengHuai |
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| Abstract: | A pair of primers was designed and synthesized according to glycoprotein D gene sequence of pseudorabies virus Fa strain, The gD gene encoding area of pseudorabies virus Fa strain exclude signal peptide was amplified by PCR from the recombinant plasmid pB13, which harbors the complete gD gene. The PCR product was cloned into an expression vector pThiohisA for E. coli and then sequenced. The sequencing results showed that gD gene encoding area exclude signal peptide is composed of 1155 bp, which is completely same with the sequence of gD gene of PRV Fa strain. The recombinant plasmid pThiohisAgD was transformed into XL1 blue, Top10,DE3 and DH5a respectively, then the expression was induced by adding IPTG to a final concentration of 1 mmol/L. SDS-PAGE result indicated the expression level was too low, and the expression level in XL1-blue was the highest. Western-blotting further confirmed glycoprotein D expressed in E. coli was antigenic. |
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| Keywords: | pseudorabies virus gD gene encoding area exclude signal peptide Escherichia coli induce expression antigenic |
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