人胃印戒细胞癌组织cDNA消减文库的构建

目的 构建人胃印戒细胞癌组织cDNA消减文库。方法 采用抑制性消减杂交技术，以胃印戒细胞癌组织为检测子，正常胃黏膜组织为驱动子，分离胃印戒细胞癌组织特异性表达的基因片段，并与T载体连接，构建人胃印戒细胞癌组织cDNA消减文库。结果 成功构建高消减效率的人胃印戒细胞癌组织cDNA消减文库，获得200多个阳性克隆，随机挑取50个克隆鉴定均有长度为100～750bp插入片段。结论应用抑制性消减杂交技术构建人胃印戒细胞癌组织cDNA消减文库，为进一步筛选和克隆人胃印戒细胞癌组织特异性表达基因奠定基础。

Construction of A Subtracted cDNA Library of Differentially Expressed Genes in Signet Ring Cell Carcinoma of the Stomach

Objective To construct a subtracted cDNA library of human genes expressed exclusively or preferentially in signet ring cell carcinoma of the stomach. Methods Suppression subtractive hybridization(SSH) was performed to isolate up-regulated cDNA fragments in signet ring cell stomach carcinoma. Human gastric signet ring cell carcinoma tissue was used as the tester, and adjacent normal stomach tissue was used as the driver. Then these cDNA fragments were directly inserted into T/A cloning vector to set up a subtraction cDNA library of specially or highly expressed genes in gastric signet ring cell carcinoma. Results More than 200 positive clones were obtained. Fifty clones were randomly picked and identified to have contained inserts in the range 100~750 bp. Conclusion A subtracted {cDNA} library of differentially expressed genes in human gastric signet ring cell carcinoma is constructed successfully with SSH and T/A clonning techniques. The library is efficient for screening and clonning specific and highly expressed genes in signet ring cell carcinoma of the stomach.;