AMY1基因编码α淀粉酶在体外培养大鼠颌下腺细胞中的表达  

The assay of the expression of α-amylase encoded by the AMY-1 gene in cultured cell of rat submandibular gland

作　　者：周青[1] 隋承光[2] 王玉新[1] 

机构地区：[1]中国医科大学附属口腔医院口腔颌面外科,辽宁沈阳110002 [2]中国医科大学附属第一医院肿瘤研究所第二研究室

出　　处：《中国医科大学学报》2007年第6期680-682,共3页Journal of China Medical University

基　　金：辽宁省科技攻关基金资助项目(00225001)

摘　　要：目的:检测大鼠颌下腺(SMG)AMY1基因编码α淀粉酶(α-Amy)在体外培养颌下腺细胞中的表达,鉴定细胞来源和检测体外培养细胞的功能。方法:分别进行培养大鼠细胞RT-PCR检测淀粉酶的基因表达;免疫印迹法(Western blotting)检测淀粉酶蛋白质。结果:大鼠SMG组织和培养细胞检测在474bp处显示有一相同的带。表明大鼠培养SMG细胞RT-PCR产物与大鼠SMG组织α-淀粉酶mRNA表达相同。免疫印迹结果显示大鼠SMG组织与培养细胞分别出现了一单独的α-淀粉酶免疫活性蛋白电泳带,其分子大小为50kDa,两者免疫活性蛋白泳带一致。结论:大鼠SMG组织和培养细胞淀粉酶mRNA表达相同,大鼠培养SMG细胞与大鼠SMG组织具有同源性。体外培养SMG细胞通过AMY1基因编码α-淀粉酶表达。Objective：To assay the expression of α-amylases genes in the cultured cells of submandibular gland（SMG）.Methods：Tissues and the cultured cells of rat submandibular gland were homogenized and crude α-amylases were extracted.The molecular weight of amylase was determined by Western blotting.The amylase gene expression of cultured cells was assayed by RT-PCR.Results：The amylase gene expressions of cultured cells and the submandibular gland cells were assayed respectively by RT-PCR.RT-PCR of total RNA was done with a specific primer for the consensus sequence of rat amylases.The same band of product at 474-bp was found in both of cultured cells and submandibular gland cells,indicating that the RT-PCR product of cultured cells was the same as mRNA of α-amylase in submandibular gland cells,and thus there was homology between the cultured cells and the cells of SMG.Amylase protein was assayed by Western blotting.The same antibody of amylase was used and the result showed that amylases of submandibular gland from rat were 50 kDa,the same as that of the cultured cells on Western blotting analysis.Conclusion：α-amylase detected in SMG tissues and cultured cells encoded by the AMY 1 gene is a SMG-specific amylase expressed in cultured cells.There is homology between SMG tissues and cultured cells,which is expressed through amylases encoded by AMY 1 gene.

关 键 词：颌下腺 细胞培养 Α-淀粉酶 

分 类 号：Q813.3[生物学—生物工程]