摘要
目的 :克隆妊高征胎盘特异表达基因并阐述它在该病发病中的意义。方法 :以正常胎盘和妊高征胎盘为模型 ,利用基于PCR的改良消减杂交技术 ,建立与妊高征相关的cDNA消减文库。随机筛选出 2 0个含有差异表达片段的克隆备用。结果 :2 0个含有差异片段的克隆经过反向点杂交 ,弃掉 6个假阳性克隆 ,确定 14个克隆为妊高征发病相关基因。经过序列测定和局部相似性检索 (BLAST) ,发现 11个为已知基因 ,其中 1号基因为NECDIN家族基因 ;3个为新基因。这 3个新基因已完成了全部插入片段的序列测定 ,并已登录GeneBank ,登录号为AF2 32 2 16、AF2 32 2 17及AF2 33648。结论 :本研究首次报道了这些基因 (尤其是Necdin表达基因 )在妊高征中高表达 ,这有助于进一步探讨妊高征的病因。
Objective:To clone genes specifically expressed in placenta of PIH,then,trying to explain their significance in the etiology of PIH.Methods:In this study,the placenta of PIH and normotention were used as model,the cDNA library was constructed and 20 fragments differentially expressed were cloned after a new version of PCR-based subtractive hybridization.Results: There were 6 false positive clones identified by reverse dot blotting,and the rest 14 clones were identified as PIH related genes.These clones were sequenced,then analyzed with BLAST analysis,and 11 of 14 clones were genes already known,and the rest 3 were identified as novel genes.These 3 genes were accepted by Gene Bank.Their accession numbers are AF232216,AF232217,AF233648. Conclusions:It is practical for cloning new genes by this strategy,and this study is the first to report these genes are related with etiology of PIH.They will be helpful in further understanding the etiology of PIH.
出处
《现代妇产科进展》
CSCD
2001年第2期83-85,共3页
Progress in Obstetrics and Gynecology
关键词
妊娠高血压综合征
胎盘
基因克隆
聚合酶链反应
减法杂交
Pregnancy induced hypertension
Placenta
Gene clone
Polymerase chain reaction
Subtractive hybridization
