戊型肝炎病毒(HEV)开读框架2中1.3 kb cDNA的克隆与序列分析
作者：蒋琳 杨恭 沈心亮
单位：蒋琳 沈心亮(卫生部兰州生物制品研究所，兰州 730046)；杨恭(兰州大学生物系，兰州 730000)
关键词：戊型肝炎病毒；cDNA克隆；测序
中国病毒学000306摘要：以戊型肝炎(Hepatitis E)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)1.3 kb基因片段，用EcoR I和BamH I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1 309 bp，其A=249(19.2%)，G=318(24.29%)，T=339(25.9%)，C=403(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%。
中图分类号：R512.36 文献标识码：A
, 百拇医药
文章编号：1003-5125(2000)03-0237-06
Cloning and Sequence Analysis of the 1.3 kb cDNA Fragment
from Open Reading Frame 2 (ORF2) of Hepatitis E Virus (HEV)
JIANG Lin，SHEN Xin-liang
(Lanzhou Institute of Biological Products, Lanzhou 730046, China)
YANG Gong
(Biological Department of Lanzhou University, Lanzhou 730000, China)
, 百拇医药
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.
, http://www.100md.com
Key word: Hepatitis E virus; cDNA cloning; Sequencing
作者简介：蒋琳(1962年-)，女，上海人，研究员，研究方向：分子生物学。
参考文献
[1] Bradley D W. Hepatitis E: Epidemiology, aetiology and molecular biology [J]. Rev Med Virol, 1992,2:19-28
[2] Tsarve S A, Emerson S U, Reyes G R, et al. Characterization of a prototype strain of Hepatitis E virus [J]. Proc Nati Acad Sci, 1992,89:559-563
, 百拇医药
[3] Favorov M O, Fields H A, Purdy M M, et al. Serologic identification of Hepatitis E virus infection in epidemic and enddemic setting [J]. J Med Virol, 1992,36:246-249
[4] Chauhau A, Jmeel S, Dilawari J B, et al. Hepatitis E virus Transmission to volunteer [J]. Lancet, 1993,341:149-150
[5] Tam A W, Smith M M, Egueraetal M. Hepatitis E virus: Molecular cloning and sequencing of the full-length viral genome [J]. Vorology, 1991,185:120-131
, 百拇医药
[6] Aye T T, Uchina T, Ma X, et al. Sequence and structure of hepatitis E virus isolated from Myanmar [J]. Virus Genes, 1993,7:95-109
[7] Aye T T, Uchina T, Ma X, et al. Complete Sequence of hepatitis E virus isolated from the Xinjiang epidemic (1986-1988) of China [J]. Nuc Acids Res, 1992,20:3512
[8] Huang CC, Nguyen D, Fernander J, et al. Molecular cloning and sequencing of the Mexico isolate of Hepatitis E virus [J]. Virology, 1992,191:550-558
, http://www.100md.com
[9] Van Cuyck-Gandre H, Zhang H Y J,et al. Characterization of Hepatitis E virus from Algeria and Chad by partial genome sequence [J]. J med Viro, 1997,53:340-347
[10] Panda S K, Nanda S K, Zafrullah M, et al. An Indian strain of hepatitis virus: cloning, sequence, and expression of structural region and antibody responses in sera from an area of high-level HEV endemicity [J]. J. Clin. Microbiol. 1995,33:2653-2659
[11] Donati M C, Fagan E A, Harrison T J, in: Rizzetto M. Purcell R.H, Gerin J L, Verme G (eds.); Viral Hepatitis and Liver Disease [M], 1997,313-316
[12] Sambrook J, Fritsch E F, Maniatis T. Molecular cloning: A laboratory manual 2nd ed. Cold Spring HarborLab Press, New York,1989
[13] Ausubel F M, Brent R, Kingston R E, et al. Short Protocols in Molecular Biology. (1995, Third Edition)
收稿日期：1999-06-28，修回日期：1999-10-13, http://www.100md.com
单位：蒋琳 沈心亮(卫生部兰州生物制品研究所，兰州 730046)；杨恭(兰州大学生物系，兰州 730000)
关键词：戊型肝炎病毒；cDNA克隆；测序
中国病毒学000306摘要：以戊型肝炎(Hepatitis E)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)1.3 kb基因片段，用EcoR I和BamH I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1 309 bp，其A=249(19.2%)，G=318(24.29%)，T=339(25.9%)，C=403(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%。
中图分类号：R512.36 文献标识码：A
, 百拇医药
文章编号：1003-5125(2000)03-0237-06
Cloning and Sequence Analysis of the 1.3 kb cDNA Fragment
from Open Reading Frame 2 (ORF2) of Hepatitis E Virus (HEV)
JIANG Lin，SHEN Xin-liang
(Lanzhou Institute of Biological Products, Lanzhou 730046, China)
YANG Gong
(Biological Department of Lanzhou University, Lanzhou 730000, China)
, 百拇医药
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.
, http://www.100md.com
Key word: Hepatitis E virus; cDNA cloning; Sequencing
作者简介：蒋琳(1962年-)，女，上海人，研究员，研究方向：分子生物学。
参考文献
[1] Bradley D W. Hepatitis E: Epidemiology, aetiology and molecular biology [J]. Rev Med Virol, 1992,2:19-28
[2] Tsarve S A, Emerson S U, Reyes G R, et al. Characterization of a prototype strain of Hepatitis E virus [J]. Proc Nati Acad Sci, 1992,89:559-563
, 百拇医药
[3] Favorov M O, Fields H A, Purdy M M, et al. Serologic identification of Hepatitis E virus infection in epidemic and enddemic setting [J]. J Med Virol, 1992,36:246-249
[4] Chauhau A, Jmeel S, Dilawari J B, et al. Hepatitis E virus Transmission to volunteer [J]. Lancet, 1993,341:149-150
[5] Tam A W, Smith M M, Egueraetal M. Hepatitis E virus: Molecular cloning and sequencing of the full-length viral genome [J]. Vorology, 1991,185:120-131
, 百拇医药
[6] Aye T T, Uchina T, Ma X, et al. Sequence and structure of hepatitis E virus isolated from Myanmar [J]. Virus Genes, 1993,7:95-109
[7] Aye T T, Uchina T, Ma X, et al. Complete Sequence of hepatitis E virus isolated from the Xinjiang epidemic (1986-1988) of China [J]. Nuc Acids Res, 1992,20:3512
[8] Huang CC, Nguyen D, Fernander J, et al. Molecular cloning and sequencing of the Mexico isolate of Hepatitis E virus [J]. Virology, 1992,191:550-558
, http://www.100md.com
[9] Van Cuyck-Gandre H, Zhang H Y J,et al. Characterization of Hepatitis E virus from Algeria and Chad by partial genome sequence [J]. J med Viro, 1997,53:340-347
[10] Panda S K, Nanda S K, Zafrullah M, et al. An Indian strain of hepatitis virus: cloning, sequence, and expression of structural region and antibody responses in sera from an area of high-level HEV endemicity [J]. J. Clin. Microbiol. 1995,33:2653-2659
[11] Donati M C, Fagan E A, Harrison T J, in: Rizzetto M. Purcell R.H, Gerin J L, Verme G (eds.); Viral Hepatitis and Liver Disease [M], 1997,313-316
[12] Sambrook J, Fritsch E F, Maniatis T. Molecular cloning: A laboratory manual 2nd ed. Cold Spring HarborLab Press, New York,1989
[13] Ausubel F M, Brent R, Kingston R E, et al. Short Protocols in Molecular Biology. (1995, Third Edition)
收稿日期：1999-06-28，修回日期：1999-10-13, http://www.100md.com