Expression of Envelope Gene gp85 of Avian Leukodls Virus in E．coli

LIU Gong-ping; ZHAO zhen-fen; LIU Fu—an

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June 2001

LIU Gong-ping, ZHAO zhen-fen and LIU Fu—an. Expression of Envelope Gene gp85 of Avian Leukodls Virus in E．coli[J]. Virologica Sinica, 2001, 16(3): 257-260.

Citation: LIU Gong-ping, ZHAO zhen-fen, LIU Fu—an. Expression of Envelope Gene gp85 of Avian Leukodls Virus in E．coli .VIROLOGICA SINICA, 2001, 16(3) : 257-260.

A亚群禽白血病病毒囊膜基因gp85片段在大肠杆菌中的表达

1.
1华南农业大学动物医学系，广东广州513640
2.
郑州牧业工程高等专科学校，河南郑州450008

摘要

将RAV-I囊膜基因 gp85片段亚克隆到表达质粒pET-21d(+)中得到重组表达质粒pET-21d-RAV-1 ca-tv( ／ sd．q，序列分析表明该插入片段的核苷酸序列和阅读框都与RAV．1囊膜基因相应序列相同。用其转化大肠杆菌 BL21(DE3)并经IPTG诱导，SDS-PAGE分析表明RAV一1囊膜基因融合蛋白表达产物约20如，与理论值相符；IPTG诱导起始时间比诱导持续时间对表达量的影响更大。
- 禽白血病病毒
- , 囊膜基因gp85
- , 亚克隆
- , 原核表达

Expression of Envelope Gene gp85 of Avian Leukodls Virus in E．coli

1.
1华南农业大学动物医学系，广东广州513640
2.
郑州牧业工程高等专科学校，河南郑州450008

Abstract

A part of the cloned fragment was excised from the recombinant with Bgl II and Sal I and subcloned into the expression vector pET-21d (+) to yield another recombinant named pET-21d-RAV-1 env (Bgl II/Sal I). The recombinant was sequenced and the result showed that the inserted fragment was identical to the counterpart in RAV-1 env gene both in nucleotide sequence and open reading frame. The E.coli BL2 (DE3) transformed with recombinant plasmid were induced with 1 mmol/L IPTG and the expression product found to be 20 kD in size on SDS-PAGE. The size of the expression product was the same as that theoretically calculated. In addition, the effects of start time and duration for IPTG induction on expression efficiency were analyzed and the result indicated that the start time for IPTG induction was more important to high expression efficiency than its duration.
- Avian leukosis virus
- , env gp85 gene
- , Subcloning
- , 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

Expression of Envelope Gene gp85 of Avian Leukodls Virus in E．coli

1. 1华南农业大学动物医学系，广东广州513640
2. 郑州牧业工程高等专科学校，河南郑州450008

Abstract: A part of the cloned fragment was excised from the recombinant with Bgl II and Sal I and subcloned into the expression vector pET-21d (+) to yield another recombinant named pET-21d-RAV-1 env (Bgl II/Sal I). The recombinant was sequenced and the result showed that the inserted fragment was identical to the counterpart in RAV-1 env gene both in nucleotide sequence and open reading frame. The E.coli BL2 (DE3) transformed with recombinant plasmid were induced with 1 mmol/L IPTG and the expression product found to be 20 kD in size on SDS-PAGE. The size of the expression product was the same as that theoretically calculated. In addition, the effects of start time and duration for IPTG induction on expression efficiency were analyzed and the result indicated that the start time for IPTG induction was more important to high expression efficiency than its duration.