Amplification and Orientation of Dengue 2 Virus Partial E Gene Insert in pUC18 by Universal Primer Directed PCR

CHEN Huo-Sheng; GUO Hui-Yu; LIU Chang-Xiu; TANG Wei-Qi; SHI Song; QIU Zhong-Ren; HUANG Xiao-Jun; ZHUNG Jian

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August 1992

CHEN Huo-Sheng, GUO Hui-Yu, LIU Chang-Xiu, TANG Wei-Qi, SHI Song, QIU Zhong-Ren, HUANG Xiao-Jun and ZHUNG Jian. Amplification and Orientation of Dengue 2 Virus Partial E Gene Insert in pUC18 by Universal Primer Directed PCR[J]. Virologica Sinica, 1992, 7(4).

Citation: CHEN Huo-Sheng, GUO Hui-Yu, LIU Chang-Xiu, TANG Wei-Qi, SHI Song, QIU Zhong-Ren, HUANG Xiao-Jun, ZHUNG Jian. Amplification and Orientation of Dengue 2 Virus Partial E Gene Insert in pUC18 by Universal Primer Directed PCR .VIROLOGICA SINICA, 1992, 7(4) : 408.

通用引物PCR分离登革病毒2型cDNA及其插入方向鉴定

1.
中山医科大学微生物学教研室
2.
中山医科大学中心实验室
3.
汕头大学医学院微生物学教研室广州

摘要

逆转录后采用聚合酶链反应(RT/PCR)扩增登革病毒2型中国海南98株(DEN_2HN98)部分包膜糖蛋白(E)基因,PCR产物钝端插入pUC18质粒,获得重组质粒pDEⅡ305。用pUC/M13测序用通用引物,PCR方法又从pDEⅡ305扩增分离DEN_2 E cDNA片段。通过一侧通用引物和另一侧DEN_2 E特异引物配对,引导酶促DNA扩增反应,鉴定了pDEⅡ305中cDNA片段的插入方向,结果与序列分析一致。本文首次报道通用引物PCR方法的建立,实验结果表明该技术可用于pUC/M13系统中插入片段的分离及其方向鉴定。
- PCR
- , 通用引物
- , 登革病毒 cDNA

Amplification and Orientation of Dengue 2 Virus Partial E Gene Insert in pUC18 by Universal Primer Directed PCR

1.
DePartment of Microbiology
2.
Sun Yatsen University of Medical Sciences
3.
Guangzhou 510089

Abstract

A portion of dengue 2 virus strain Haina 98 (DEN2 HN98)envelope (E1-305 nt) gene has been isolated by reverse transcription and polymerase chain reaction(RT-PCR). The plasmid pDEⅡ305 was constructed by blunt ligation of the PCR products of 305 bp fragment with SmalⅠdigested pUC18. The DEN2 E305 insert were isolated and amplified from pDEⅡ305 by universal primer directed PCRo The pUC/M13 sequencing primers (forward and reverse, Promega)and DEN2 E.specific primers(forward and reverse, Promege) and DEN2 E spec...
- PCR UniversaI Primer Dengue. Virus cDNA

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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

Amplification and Orientation of Dengue 2 Virus Partial E Gene Insert in pUC18 by Universal Primer Directed PCR

1. DePartment of Microbiology
2. Sun Yatsen University of Medical Sciences
3. Guangzhou 510089

Abstract: A portion of dengue 2 virus strain Haina 98 (DEN2 HN98)envelope (E1-305 nt) gene has been isolated by reverse transcription and polymerase chain reaction(RT-PCR). The plasmid pDEⅡ305 was constructed by blunt ligation of the PCR products of 305 bp fragment with SmalⅠdigested pUC18. The DEN2 E305 insert were isolated and amplified from pDEⅡ305 by universal primer directed PCRo The pUC/M13 sequencing primers (forward and reverse, Promega)and DEN2 E.specific primers(forward and reverse, Promege) and DEN2 E spec...