摘要
根据国外报道的鸡 IL- 1β序列 ,设计引物 ,以提取的鸡外周血单个核细胞总 RNA为模板 ,RT- PCR扩增 IL- 1β c DNA全序列。将 RT-PCR产物与 p UC19载体相连 ,转化大肠杆菌 ,经 PCR、酶切分析及序列测定得知所克隆序列与报道序列同源性达 99%以上 ,证明所克隆基因为目的基因。用地高辛随机引物法标记 IL- 1βc DNA制备探针 ,原位杂交检测禽脑脊髓炎病鸡与正常鸡脑组织中 IL- 1β m RNA表达情况 ,结果表明脑炎病鸡脑中 IL - 1β m RNA表达水平高于正常鸡。
According to the described sequence of chicken interleukin 1β(ChIL 1β) and by using the extracted total RNA of chicken peripheral blood mononuclear cell (PMBC), a pair of primers were designed to reverse transcriptionally amplifying ChIL 1β gene. Connecting the production of RT PCR with the plasmid vector of pUC19 and transforming the vector into E.coli,the recombint bodies of ChIL 1β were produced. Then they were examined by PCR amplifying,enzyme digesting and sequence analysis. Analysis of ChIL 1β showed that it had more than 99% homology identity to the described ChIL 1β sequence.The double chains of ChIL 1β cDNA were labeled with Dig 11 dUTP to produce probe.And the ChIL 1β mRNA secreation was studied in the tissues of chickens' brains infected with AEV and that of normal chickens'. The results showed that the content of ChIL 1β secreation in sick chickens' brain was higher than that of the normal chickens'
出处
《动物医学进展》
CSCD
2003年第3期85-87,共3页
Progress In Veterinary Medicine
基金
国家自然科学基金资助项目(39960 0 61 )
