两种来源的鲑鱼降钙素基因的序列分析与比较  

SEQUENCING AND COMPARING OF TWO SALMON CALCITONIN GENES FROM DIFFERENT SOURCES

作　　者：蔡芳[1] 邓彤[1] 韩剑鸣 张镜宇[1] 

机构地区：[1]天津医科大学内分泌研究所

出　　处：《天津医科大学学报》1999年第1期1-4,共4页Journal of Tianjin Medical University

基　　金：天津市自然科学基金

摘　　要：目的：为明确鲑鱼降钙素基因的ＤＮＡ结构，克隆鲑鱼降钙素基因（ｓＣＴ）并经序列分析予以确证。方法：分别从市售进口常用试剂鲑精ＤＮＡ和自我国东北鲑鱼新鲜分离的精巢组织ＤＮＡ中，利用聚合酶链反应，分离出鲑鱼降钙素基因的编码序列，并分别将其插入克隆载体ｐＵＣ１８，经双脱氧末端终止法进行序列分析。结果：证实所得两种基因克隆均含编码ｓＣＴ降钙素３２个氨基酸的全长ＤＮＡ序列９６ｂｐ，二者无差异。结论：已获得鲑鱼降钙素基因克隆。Objective: For determining the DNA structure of salmon calcitonin(sCT) gene, the necessary step is to clone and sequence it Methods:Owing to the location of the encoding DNA fragment of sCT 1 32 in the same extron, and different reports about sCT gene's DNA structure, two sCT genes were separated directly from commercial salmon sperm DNA(GIBCO) and salmon(northeast, China) testis tissue DNA respectively by means of PCR method, and identified by 2% agrose gel electrophoresis or 10% polyacrylamide gel electrophoresis using silver stain(Figure 1,2) The PCR products were inserted between the SmaⅠ and EcoRⅠ sites of pUC18 vector respectively The nucleotide sequence of the DNA fragments was determined with the dideoxy chain termination method combining with PCR technique Results: Two salmon calcitonin genes from different sources were cloned and shown no difference of their DNA structure, both of which encode the 32 amino acids of sCT as reported before Conclusion: Salmon calcitonin genes from two different sources were sequenced and compared, which was a preliminary work of producing sCT polypeptide

关 键 词：鲑鱼 降钙素基因 聚合酶链反应 序列分析 

分 类 号：Q51[生物学—生物化学]