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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 transcriptional status of Bam HIL fragment of virulent GA strain of Marek ’s disease (MDV) in lymphoid tissue induced by MDV virulent strain Jing - 1 (endemic to China). Methods m RNAs were isolated from visceral lymphatic neoplasms of chickens infected with MDV-induced virulence strain Jing-1. Based on the published m eq gene sequences of MDV tumor candidate genes and the Bam HIL fragment sequences of virulent GA strains that we determined, Two oligonucleotide primers, m RNA as a template by reverse transcription of cDNA synthesis, PCR method for gene amplification. Genomic Bam HII2 and LDNA fragments of GA strain were labeled with non-radioactive Digoxigenin to generate nucleic acid probe, which was respectively identified by Southern blot hybridization with PCR products. Results The size of cDNA amplified by reverse transcriptase-polymerase chain reaction (RT-PCR) was about 730 bp. The cDNA was able to react with the above two probes simultaneously in immunochromism. Conclusion ①MVDV m eq gene transcription can be extended to the right Bam HIL fragment; ② Bam HIL region can be transcribed in the MDV tumorigenic Jing-1 strain induced lymphoma.