β3GnT2、β3GnT8真核表达载体的构建及胃癌AGS、胶质瘤U251细胞株中β3GnT8cDNASNP的分布

摘要

Objective Cloning of B 3GnT2 and β 3GnT8 and construction of their eukaryotic expression vectors.Methods RT-PCR was used to detect β 3GnT2, β 3GnT8 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 pEGFPC1- β 3GnT2,pEGFP-C1- β 3GnT8 were constructed.Results β 3GnT2, β 3GnT8 were expressed in all cell lines shown above;The whole encoding fragments of B 3GnT2, β 3GnT8 with restriction endonuclease recognition sites were constructed into pEGFP-C1 vector;there are four β 3GnT8 cDNA SNP sites both in Gastric cancer AGS and glioma U251 cell lines.Conclusion β 3GnT2, B 3GnT8 were both expressed in all cell lines detection,the whole encoding fragments of β 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.%目的 β3GnT2和β3GnT8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pEGFP-C1-β3GnT2、pEGFP-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U251细胞株中β3GnT8 cDNA经测序鉴定存在4个一致的SNP位点.结论 β3GnT2、β3GnT8共同表达于多种癌细胞株中;成功克隆β3GnT2、β3GnT8全长编码片段并构建真核表达载体pEGFP-C1-β3GnT2、pEGFP-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.