Cloning and Sequence Analysis of the 1.3 kb eDNA Fragment from Open Reading Frame 2(ORF2)of Hepatitis E Virus(HEV)

JIANG Lin; YANG Gong; CHEN Xin-Liang

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June 2000

JIANG Lin, YANG Gong and CHEN Xin-Liang. Cloning and Sequence Analysis of the 1.3 kb eDNA Fragment from Open Reading Frame 2(ORF2)of Hepatitis E Virus(HEV)[J]. Virologica Sinica, 2000, 15(3): 237-242.

Citation: JIANG Lin, YANG Gong, CHEN Xin-Liang. Cloning and Sequence Analysis of the 1.3 kb eDNA Fragment from Open Reading Frame 2(ORF2)of Hepatitis E Virus(HEV) .VIROLOGICA SINICA, 2000, 15(3) : 237-242.

戊型肝炎病毒(HEV)开读框架2中1.3kbcDNA的克隆与序列分析

1.
卫生部兰州生物制品研究所，兰州730046

摘要

以戊型肝炎 (HepatitisE)病人高热期血清为原材料 ,设计特定引物 ,经RT PCR扩增获得开放阅读框 2 (ORF2 ) 1.3kb基因片段 ,用EcoRI和BamHI插入克隆载体 pUC18,测定了该克隆基因片段的核苷酸序列并进行了比较分析。其片段长度为 130 9bp ,其A =2 4 9(19.2 % ) ,G =318(2 4 .2 9% ) ,T =339(2 5.9% ) ,C =4 0 3(30 .79% ) ;与印度株和缅甸株的同源性在 92 %以上 ,而与乍得和墨西哥株的同源性分别为 90 .7%和 77.4 %。
- 戊型肝炎病毒
- , cDNA克隆
- , 测序

Cloning and Sequence Analysis of the 1.3 kb eDNA Fragment from Open Reading Frame 2(ORF2)of Hepatitis E Virus(HEV)

1.
Lanzhou

Abstract

The serum from patients infected with Hepatitis E was prepared, the RNA of Hepatitis E virus was purified. The interested cDNA fragment (1.3?kb) was obtained by RT PCR, cloned in to the plasmid pUC18 and sequenced by the dideoxynucleotide method (fluorescein labeled dNTP). The result of sequencing suggested that the cDNA fragment was 1?309?bp, its adenine, guanine, thymine and cytosine were 249 (19.2%), 318(24.29%), 339(25.9%) and 403(30.79%), respectively. The nucleic acid sequences of the cDNA were 92% to 93.3% homologous to the Indian and Burmese strains, 90.7% and 77.4% homologous to Chad and Mexican strains. The ORF2 of HEV is relatively stable between the Asian strains.
- Hepatitis E virus
- , cDNA cloning
- , Sequencing

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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 Sequence Analysis of the 1.3 kb eDNA Fragment from Open Reading Frame 2(ORF2)of Hepatitis E Virus(HEV)

1. Lanzhou

Abstract: The serum from patients infected with Hepatitis E was prepared, the RNA of Hepatitis E virus was purified. The interested cDNA fragment (1.3?kb) was obtained by RT PCR, cloned in to the plasmid pUC18 and sequenced by the dideoxynucleotide method (fluorescein labeled dNTP). The result of sequencing suggested that the cDNA fragment was 1?309?bp, its adenine, guanine, thymine and cytosine were 249 (19.2%), 318(24.29%), 339(25.9%) and 403(30.79%), respectively. The nucleic acid sequences of the cDNA were 92% to 93.3% homologous to the Indian and Burmese strains, 90.7% and 77.4% homologous to Chad and Mexican strains. The ORF2 of HEV is relatively stable between the Asian strains.