马立克病疱疹病毒致瘤株基因组IRL区Bam H I I2和L片段内一特异性cDNA的分离与鉴定  

Isolation and Identification of a Specific cDNA Mapping to the BamHI I2 and L Fragments within the Inverted Repeats of Unique Long Region (IRL) in the Genome of Marek′s Disease Herpesvirus (MDV) Oncogenic Strain Beijing 1

作　　者：卢春[1] 吴建平[2] 张训海[3] 朱鸿飞[3] 蔡宝祥[3] 

机构地区：[1]南京医科大学微生物学与免疫学教研室 [2]南京医科大学毒理学研究所,南京210029 [3]南京农业大学动物医学院畜禽传染病学实验室,南京210095

出　　处：《南京医科大学学报（自然科学版）》1999年第6期447-449,共3页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家自然科学基金

摘　　要：目的　了解马立克病疱疹病毒（ＭＤＶ）强毒ＧＡ株Ｂａｍ ＨＩＬ片段在ＭＤＶ致瘤毒株京－１株（中国特有的地方株）所诱导的淋巴肿瘤组织中转录状况。方法　从人工感染ＭＤＶ致瘤株京－１株发病鸡内脏淋巴肿瘤组织中分离ｍ ＲＮＡ，根据已发表的ＭＤＶ肿瘤候选基因ｍ ｅｑ 基因序列和我们已测定的强毒ＧＡ株Ｂａｍ ＨＩＬ片段序列，设计两段寡核苷酸引物，以此ｍ ＲＮＡ为模板经逆转录合成ｃＤＮＡ，应用ＰＣＲ法进行目的基因扩增。用非放射性Ｄｉｇｏｘｉｇｅｎｉｎ 分别标记ＧＡ株基因组Ｂａｍ ＨＩＩ２和ＬＤＮＡ片段，制成核酸探针，分别与ＰＣＲ产物作Ｓｏｕｔｈｅｒｎ ｂｌｏｔ分子杂交鉴定。结果　逆转录聚合酶链反应（ＲＴ－ＰＣＲ）扩增的ｃＤＮＡ大小约７３０ ｂｐ，该ｃＤＮＡ能同时与上述两探针发生免疫呈色反应。结论　①ＭＤＶ ｍ ｅｑ基因的转录可以延伸到其右侧的Ｂａｍ ＨＩＬ片段区；②Ｂａｍ ＨＩＬ区在ＭＤＶ致瘤株京－１株所诱导的淋巴肿瘤组织中可发生转录。Objective\ To understand the transcription of BamHI L DNA fragment from genome of strong virulent GA strain of Marek′s disease herpesvirus (MDV) in lymphoblastoid tumor tissue induced by oncogenic strain Beijing 1 (a specific local strain in China) of MDV. Methods\ Two oligonucleotide primers were synthesized according to the reported sequence of \%meq\% gene an ideal oncogenic candidate and our previously determined sequence of BamHI L fragment of Marek′s disease herpesvirus (MDV), respectively. Reverse transcriptase PCR(RT PCR) assay was performed by using these primers and the mRNA as a template which was isolated from visceral lymphoblastoid tumors obtained from chickens artificially infected with strain Beijing 1 of oncogenic MDV. Southern blot molecular hybridization was further carried out to detect the product of RT PCR with digoxigenin labeled nucleotide probe from BamHI I2 and L fragment in the gene library of MDV strain GA, respectively. Results\ Two probes could simultaneously hybridize this cDNA amplified by RT PCR with a length of about 730 bp. Conclusion\ It is suggested that \%meq\% transcription could extend from the right hand end of BamHI I2 to the adjacent BamHI L, and the BamHI L region was likely to be transcribed in MDV induced lymphoblastoid tumors.

关 键 词：疱疹病毒 MEQ基因 BamHIL片段 基因转录 MDV 

分 类 号：R373.11[医药卫生—病原生物学]