Cloning and Determ ination of S 1 Gene of IBV GD05 Strain and Its Expression in E ．coli．

HUANG Ya; ZHEN Qing; WANG Lin; LI Xiao-kun; HUANG Zi

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June 2002

HUANG Ya, ZHEN Qing, WANG Lin, LI Xiao-kun and HUANG Zi. Cloning and Determ ination of S 1 Gene of IBV GD05 Strain and Its Expression in E ．coli．[J]. Virologica Sinica, 2002, 17(3): 266-269.

Citation: HUANG Ya, ZHEN Qing, WANG Lin, LI Xiao-kun, HUANG Zi. Cloning and Determ ination of S 1 Gene of IBV GD05 Strain and Its Expression in E ．coli． .VIROLOGICA SINICA, 2002, 17(3) : 266-269.

IBV广东分离株GD05 S1基因的克隆、鉴定及其表达

1.
1．暨南大学医药生物技术研究开发中心，广州510632
2.
华南农业大学动物医学系，广州510642
3.
华南农业大学蚕桑系．广州510642

摘要

Acording tO Avian Infectious Brochitis virus(IBV)Beaudette strain S 1 gene sequence，a pair of primers were designed and synthesized．W ith the primers， IBV Guangdong isolation strain GD05 S 1 gene was successively amplified by RT — PCR ．PCR product was digested w ith BstY I，Hae III and Pst I respectively，the result showed RFLP pattern of was the same as that of M41 S 1 gene． IBV GD05 strain was thought as M ass serotype primaryly．IBV GD05 S 1 gene was cloned into pGEM — T vector and sequenced， its sequence was consisted of 1611 base pairs．By comparison ， the nu— cleotide sequence was 97．14％ identical tO that of IBV M 41． IBV GD05 S 1 gene was subcloned into expression vector pET21 d．SDS—PAGE experiment showed that it expresed in E ．coli．
- IBV
- , S1基因
- , 克隆
- , 表达

Cloning and Determ ination of S 1 Gene of IBV GD05 Strain and Its Expression in E ．coli．

1.
1．Biopharmaceutic R&D center of Jinan University，Guangzhou 510632，China
2.
Department of veterinary medicine South china Agriculture University，@uangzhou 510642，China
3.
Depa rtment of Sericulture， South china Ag riculture University，Guangzhou 510642，China

Abstract

Acording tO Avian Infectious Brochitis virus(IBV)Beaudette strain S 1 gene sequence，a pair of primers were designed and synthesized．W ith the primers， IBV Guangdong isolation strain GD05 S 1 gene was successively amplified by RT — PCR ．PCR product was digested w ith BstY I，Hae III and Pst I respectively，the result showed RFLP pattern of was the same as that of M41 S 1 gene． IBV GD05 strain was thought as M ass serotype primaryly．IBV GD05 S 1 gene was cloned into pGEM — T vector and sequenced， its sequence was consisted of 1611 base pairs．By comparison ， the nu— cleotide sequence was 97．14％ identical tO that of IBV M 41． IBV GD05 S 1 gene was subcloned into expression vector pET21 d．SDS—PAGE experiment showed that it expresed in E ．coli．
- Avian Infectious Bronchitis Virus(IBV)
- , S 1 gene
- , Cloning
- , Expression

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Cloning and Determ ination of S 1 Gene of IBV GD05 Strain and Its Expression in E ．coli．

1. 1．Biopharmaceutic R&D center of Jinan University，Guangzhou 510632，China
2. Department of veterinary medicine South china Agriculture University，@uangzhou 510642，China
3. Depa rtment of Sericulture， South china Ag riculture University，Guangzhou 510642，China

Abstract: Acording tO Avian Infectious Brochitis virus(IBV)Beaudette strain S 1 gene sequence，a pair of primers were designed and synthesized．W ith the primers， IBV Guangdong isolation strain GD05 S 1 gene was successively amplified by RT — PCR ．PCR product was digested w ith BstY I，Hae III and Pst I respectively，the result showed RFLP pattern of was the same as that of M41 S 1 gene． IBV GD05 strain was thought as M ass serotype primaryly．IBV GD05 S 1 gene was cloned into pGEM — T vector and sequenced， its sequence was consisted of 1611 base pairs．By comparison ， the nu— cleotide sequence was 97．14％ identical tO that of IBV M 41． IBV GD05 S 1 gene was subcloned into expression vector pET21 d．SDS—PAGE experiment showed that it expresed in E ．coli．