合成人降钙素基因实验研究的初步报告

目的 :合成含人胰岛素分泌信号的人降钙素基因。方法 :将人胰岛素分泌信号和人降钙素基因的序列分成六个片断合成单链寡核苷酸作为PCR的引物和cDNA合成的模板。应用动态模板PCR反应直接扩增出人胰岛素和降钙素的编码序列。结果 :经含EB的 1.5 %琼脂糖凝胶电泳 ,证明合成的基因含编码人胰岛素分泌信号和人降钙素的全长DNA序列。结论 :本研究为基因合成提供了一种新的可靠方法。

Preliminary study of human calcitonin gene synthesis

Objective: To recombine the gene of human calcitonin (hCT) which consists the insulin secretory signal peptide (ISSP)Methords:A gene coding for human calcitonin and insulin secretory signal peptide was divided into six fragments, synthesis by changing template PCR technique.The 212bp hCT and ISSP gene synthesis does not rely on the DNA ligase but relies on fidelity DNA polymerase LA TaqTM . The hCT and ISSP gene was assembled through three round of PCR from a total of only six 40-60bp oligos.Results:Finally,the nucleotide sequence of synthesized hCT and ISSP gene was determined with the method of agarglucose electrophoresis.The result indicates that the nucleotide sequence of sequence of the hCT and ISSP gene is the same as the sequence designed. Conclusion:The result demonstrates that a reliable and forthright method of synthesis of oligo-peptide gene (or cDNA) is provided.This paper is a basic work for expression and secretion of hCT ,or further study on hCT-associated disease.