人白细胞介素29的cDNA克隆和序列分析  

Cloning and Sequencing of Human IL-29 cDNA

作　　者：施国民[1] 张锦海[1] 李越希[1] 吴玉章[2] 

机构地区：[1]南京军区军事医学研究所,南京210002 [2]第三军医大学免疫学教研室,重庆400038

出　　处：《中国生物工程杂志》2004年第10期34-37,共4页China Biotechnology

摘　　要：目的 :克隆人白细胞介素 2 9的cDNA ,以进一步研究其结构与功能的关系。方法 :分离人的外周血单个核细胞 ,IMDM(含PHA和IL 2 )培养过夜后 ,Poly(I:C)诱导 2 4～ 48h ,收集细胞 ,Trizol法提取细胞总RNA ,RT PCR扩增出全长 60 3bp的IL 2 9cDNA ,将其与表达载体pPROEXTMHTa连接 ,转入大肠杆菌BL2 1 (DE3) ,建立了hIL 2 9的cDNA克隆。结果 :测序表明克隆的cDNA与Genebank报道的人IL 2 9cDNA序列完全一致。结论在国内成功获得hIL 2Objective:To obtain the cDNA encoding human interleukin 29 and investigate the relationship between structure and function of hIL 29.Methods: Freshly isolated human peripheral blood mononuclear cells were grown in the presence of poly(I:C),After a 24～48 hours incubation,total RNA was used as template for RT PCR.A 603bp cDNA fragment was amplified by RT PCR,with digestion of Bam H I and Eco R I and gel extraction,the cDNA product was inserted into pPROEX TM HTa vector and transformed into E.coli /BL(DE3).The recombinant pPROEX TM HTa/hIL 29 was purified and sequenced according to the manipulation instructions.Results: After RT PCR,a specific fragment about 603bp was observed in 1% agarose gel electrophoresis,while no corresponding fragment in the negative control was found. The result of sequencing was completely consistent with that of the published hIL 29 cDNA.Conclusion: hIL 29 cDNA has been cloned successfully.

关 键 词：HIL-2 人白细胞介素2 CDNA克隆 外周血单个核细胞 HT RT-PCR 诱导 结构与功能 表达载体 CDNA序列 

分 类 号：Q785[生物学—分子生物学] R73-362[医药卫生—肿瘤]