IFITM1克隆测序及生物信息学分析  

The Cloning and Sequence Analysis of Interferon Induced Transmembrane Protein 1 Gene

作　　者：刘宇虎[1] 钟东[2] 柳娟[1] 杨兴龙[2] 张振书[2] 肖冰[2] 郭文英[1] 

机构地区：[1]广东省东莞市人民医院消化科,523018 [2]南方医科大学南方医院消化病研究所,广州510515

出　　处：《医学研究杂志》2009年第5期40-43,138,共5页Journal of Medical Research

基　　金：国家自然科学基金项目(30171053);广东省医学科学技术研究基金项目(B2005089);东莞市科技计划项目(B200501)

摘　　要：目的研究干扰素诱导的跨膜蛋白-1基因（interferon induced transmembrane protein 1．IFITM1）克隆、测序，对序列进行分析，并获取生物学信息。方法以IFITM1的cDNA为模板，用Pfu酶做PCR扩增，在EcoR Ⅰ和Hind Ⅲ双酶切后，将IF—ITM1的cDNA片段克隆到pUCm—T质粒，对重组的pUCm—T-IFITM1测序后，采用All GenBank＋EMBL＋DDBJ＋PDBSe—quences Database和Simple Modular Architecture Research Tool（SMART）对IFITM1的eDNA进行生物信息学分析。结果扩增后，含有IFITM1基因eDNA的PCR产物约1000bp，IFITM1的eDNA成功克隆到pUCm—T质粒并进行测序。序列分析结果显示，IFITM1结构cDNA全长683bp，有2个外显子。IFITM1的mRNA编码的氨基酸在37～57和87～107有2个跨膜区，编码125个氨基酸的多肽，其蛋白相对分子质量为14kDa。结论，IFITM1基因的成功扩增、克隆、测序，获取了IFITM1基因的eDNA、mRNA以及编码蛋白质氨基酸的生物学信息，有利于研究IFITM1基因与大肠癌的关系。Objective To study the cloning and sequence analysis of the interferon induced transmembrane protein 1 （IF1TM1） gene, then to obtain the biological information of IFITM1. Methods The IFITM1 gene cDNA was amplified using the Pfu enzyme by means of PCR method. After EcoR Ⅰ and Hind Ⅲ digestion, the IFITM1 gent cDNA fragment was cloned in pUCm - T plasmid and sequenced. The sequence of the IFITM1 cDNA was analyzed with All GenBank ＋ EMBL ＋ DDBJ ＋ PDB Sequences Database and Simple Modular Architecture Research Tool （SMART）. Results The length of the PCR product of IFITM1 eDNA fragment was about 1000 bp. The IFITM1 gene eDNA was successfully inserted into pUCm - T plasmid. The results of sequence analysis of the IFITM1 showed that the length of the IFITM1 cDNA was 683bp. There is two cxtrons in the sequence. IFITM1 mRNA coding protein is polypeptide of 125 amino acids,there were two transmembrane domains in amino acids sequence 37 -57 and 87 - 107. The protein molecular mass is 14kDa. Conclusion IFITM1 gene cDNA was successfully cloned, the sequence of IFITM1 cDNA was analyzed, We have obtained much biological information of the IFITM1 cDNA and mRNA and amino acids of coded protein. It was helpfully to further study on the role of IFITM1 gene in colorectal cancer.

关 键 词：结直肠癌 干扰素诱导的跨膜蛋白-1( IFITM1) 基因克隆 序列分析 生物信息 

分 类 号：R996.3[医药卫生—毒理学] R318.04[医药卫生—药学]