β3GnT2、β3GnT8真核表达载体的构建及胃癌AGS、胶质瘤U251细胞株中β3GnT8 cDNA SNP的分布  

ZHAO Shan-cheng, HU Shui-jun, ZO U Shi-tao, ZHAO Jun-yu, HAN hong-wei, WU Shi-liang （Department of Biochemistry and Molecular Biology, School ofMedicine,Soochow University, Suzhou,Jiangsu,215123,China）

作　　者：赵善成[1] 胡水君[1] 邹士涛[1] 赵俊宇[1] 韩洪伟[1] 吴士良[1] 

机构地区：[1]苏州大学医学部生物化学与分子生物学系,江苏苏州215123

出　　处：《中国血液流变学杂志》2011年第2期199-202,243,共5页Chinese Journal of Hemorheology

基　　金：国家自然科学基金资助项目（30670462）

摘　　要：目的β3GriT2和β3GriT8的克隆及真核表达载体的构建。方法RT-PCR法检测β3GnT2、β3GnT8在胃癌、胶质瘤、乳腺癌三类癌细胞株中的表达；PCR扩增带酶切位点β3GnT2、β3GnT8全长编码片段并构建真核表达载体βEGFP-C1-B3GnT2，βEGFP-C1-β3GnT8。结果β3GnT2、β3GnT8在上述癌细胞株中均有表达；带酶切位点β3GnT2、β3GnT8全长编码片段成功扩增并连接至pEGFP-C1载体上；胃癌AGS、胶质瘤U25l细胞株中β3GnT8 cDNA经测序鉴定存在4个一致的SNP位点。结论母βGnT2、β3GnT8共同表达于多种癌细胞株中；成功克隆β3GnT2、β3GnY8全长编码片段并构建真核表达载体βEGFP-C1-β3GnT2、βEGFP-C1-β3GnT8；发现胃癌AGS、胶质瘤U251细胞株中β3GnT8 cDNA SNP位点4个，为进一步的功能研究奠定了基础。Objective Cloning of β 3GnT2 and 13 3GnT8 and construction of their eukaryotic expression vectors. Methods RT-PCR was used to detect 13 3GnT2, β 3GriT8 mRNA expression level in three types of cancer cell lines including gastric,glioma and breast cancer;the whole encoding fragments of β 3GnT2, β 3GnT8 with restriction endonuclease recognition sites were amplified by PCR and their eukaryotic expression vectors pEGFPC l- β 3GnT2,pEGFP-CI- β 3GnT8 were constructed.Results β3GnT2, β 3GnT8 were expressed in all cell lines shown above;The whole encoding fragments of β3GnT2, β3GnT8 with restriction endonuclease recognition sites were constructed into pEGFP-C 1 vector;there are four β 3GnT8 cDNA SNP sites both in Gastric cancer AGS and glioma U251 cell lines. Conclusion β 3GnT2, β 3GnT8 were both expressed in all cell lines detection,the whole encoding fragments of 13 3GnT2, β 3GnT8 were cloned and their eukaryotic expression vectors were constructed successfully, and four β 3GnT8 cDNA SNP sites both in Gastric cancer AGS and glioma U251 cell lines were founded,so foundation for more extensive functional research was established.

关 键 词：β3GnT2 β 3GnT8 癌症转移 真核表达载体 单核苷酸多态性 

分 类 号：R73-3[医药卫生—肿瘤]