卵巢癌相关基因的克隆

目的：筛选和克隆卵巢上皮癌相关基因。方法：用3种锚定引种和8种随机引物，共24种组合进行PCR扩增，通过DD－RTPCR分析正常卵巢上皮和卵巢上皮癌mRNA的基因表达差异，将差异片段分离出来，并回收，克隆和进行斑点杂交鉴定、DNA序列测定，并通过国际互联网张检索GenBank进行同源性分析。结果：获得30个差异表达的片段，经斑点杂交初步筛选，克隆11个差显示的阳性片段。选取其中的5个进行序列分析。结论计算机同源性分析表明，5个均为新基因片段，已收录GenBank。

IDENTIFYING DIFFERENTIAL EXPRESSION GENESIN OVARIAN CANCER USING mRNA DIFFERENTIAL DISPLAY

Objective To identify genes related to the development of ovarian cancer by studying the differential expression genes between normal ovarian epithelia and ovary epithelia cancer. Methods The RNA of both the normal ovary epithelia and ovarian cancer cell line(3AO,SKOV3) was isolated for differentially expressed genes using differential display RT-PCR(DD-RTPCR).Three sets of anchor primers and 8 sets of arbitrary primers,total 24 primer combinations were used for DD-RTPCR amplification.These differentially expressed fragments were recovered,cloned,further analyzed by dot blot,and then sequenced these clones and compared with GenBank/EMBL databases. Results After 30 differential display bands preliminarily screened by dot blot hybridization,11 differential display fragments were cloned and 5 differential expressed fragments were sequenced. Conclusion Sequencing and homology analysis indicated that these cDNA-fragments did not match any genes published previously,they are novel gene sequences,which have been submitted to GenBank.;