人染色体17q12区雨330kb范围内表达顺序的筛选  

Screening for coding sequences in a 330 kb region on the human chromosome 17q12

作　　者：朱光旦[1] 余龙[2] 黄昕 黄浩杰 郑其平 赵寿元 

机构地区：[1]复旦大学微生物和微生物工程系 [2]复旦大学遗传学研究所,上海200433

出　　处：《复旦学报（自然科学版）》1997年第2期234-238,共5页Journal of Fudan University：Natural Science

基　　金：国家杰出青年基金;国家科委863高科技资助;国家自然科学基金;国家教委霍英东青年教授基金

摘　　要：用定位于人染色体17q12区带的长约330kbYAC克隆500D09（取自法国人类多态性研究中心YAC库）作为杂交探针，筛选人骨髓、胎脑、胎肾、骨骼肌和睾丸等五种组织的cDNA库（2×105～3×105pfu／库），从中获得102个初级阳性克隆．初级克隆经PCR扩增，分别与人基因组DNA、酵母基因组DNA和人rDNA探针作dotblot杂交分析，排除其中假阳性克隆后，复筛得到32个候选克隆．对其中2个候选克隆B4511和S5511分别测定143bp和147bp序列．经查新和同源性分析，这两个片段与已知基因的同源性均小于50％，提示它们可能是来自于新基因的表达顺序．Positional cloning of coding sequences in a 330 kb region on the human chromosome 17q12 was carried out by the direct screening method using a yeast artificial chromosome (YAC)probe. YAC 500D09 (from Centre d Etude du Polymorphisme Humain,France)located on chromosome 17q12 was used as the probe for screaning cDNA libraries of human bone marrow,fetal brain,fetal kidney,skeletal muscle and testis. A total of 102 possible positive clones were selected from these cDNA libraries with 2×105~3×105 clones. After PCR-amplified products of these clones were hybridized in dot blot with human genomic DNA,yeast genomic DNA and human rDNA probes respectively, 32 clones of them were identified as the candidate expressional sequences from YAC 500D09 and further isolated by a second round of hybridization screening. Partial sequences were obtained from two of them (B4511 and 55511) and were of the length with 143 bp and 147 bp respectively. The homology of these two were less than 50% with that of known genes,and thus we suggest that these were novel.

关 键 词：17q12区带 基因表达 遗传性疾病 染色体 

分 类 号：R596[医药卫生—内科学]