家族性进行性色素过度沉着症家系丝氨酸/苏氨酸蛋白激酶11基因的克隆和序列鉴定  

Cloning and sequence analysis of the serine/threonine protein kinase 11 in the familial progressive hyperpigmentation

作　　者：张彩娥[1] 邓云华[2] 吴雄文[3] 

机构地区：[1]华中科技大学同济医学院附属同济医院麻醉科,湖北武汉430030 [2]华中科技大学同济医学院附属同济医院皮肤科,湖北武汉430030 [3]华中科技大学同济医学院基础部免疫学教研室,湖北武汉430030

出　　处：《新乡医学院学报》2011年第5期548-550,共3页Journal of Xinxiang Medical University

基　　金：国家自然科学基金面上项目(编号:81071357);教育部博士点新教师基金资助项目(编号:200804871043)

摘　　要：目的进行家族性进行性色素过度沉着症(FPH)家系中患者丝氨酸/苏氨酸蛋白激酶11(STK11)基因的克隆和序列鉴定,以期检测FPH发生是否由于STK11突变所致。方法从FPH患者永生化的B淋巴细胞中提取总RNA,反转录聚合酶链式反应(RT-PCR)获取cDNA序列,加入特异性引物经PCR扩增STK11 cDNA双链,再经过HindⅢ、EcoRI双酶切PCR产物及质粒载体pcDNA3.0,酶切产物经回收、连接,产物转化到大肠杆菌DH5α,选取阳性克隆菌落进行PCR鉴定、酶切鉴定和测序分析。结果成功构建重组质粒pcDNA3.0-STK11,其测序结果与美国国立生物技术信息中心查询的正常序列结果一致。结论该FPH家系发病并非由于STK11突变所致,在初步定位FPH致病基因的19p13.1-pter区段存在尚未发现的致病基因。Objective To clone serine/threonine protein kinase 11（STK11） in the familial progressive hyperpigmentation（FPH） family and to analyze the mutation of STK11 relation to the cause of FPH. Methods Total RNA was extracted from cells in the FPH family for reverse transcription of the cDNA.PCR with specific primers was performed to amplify STK11 cDNA double stranded,the PCR products and plasmid vector pcDNA3.0 were cut by double-enzyme deavage with HindⅢ and EcoRI.The enzyme cut produts were recovered and jointed to target gene and vector.The recombinant plasmid pcDNA3.0-STK11 was transformed into E.coli DH5α.The positive clones were screened and their double-stranded DNA sequenced. Results Recombinant plasmid pcDNA3.0-STK11 was builded successfully.The constructed STK11 sequences were completely coincidence with GenBank. Conclusion The cause of FPH is no relation to the mutation of STK11.There is a new gene not found in restricted region for FPH.

关 键 词：家族性进行性色素过度沉着症 丝氨酸/苏氨酸蛋白激酶11基因 克隆 序列鉴定 

分 类 号：Q785[生物学—分子生物学]