RT-RAPD技术分析高温诱导双孢蘑菇相关基因片段  

Analysis of Agaricus bisporus Themotolerance Related Gene with RT-RAPD

作　　者：陈兰芬[1] 宋思扬[1] 郑忠辉[1] 苏文金[1] 陈美元[2] 廖健华 王泽生[2] 

机构地区：[1]厦门大学生命科学学院,细胞生物学与肿瘤细胞工程教育部重点实验室,福建厦门361005 [2]福建省轻工业研究所,福建福州350005

出　　处：《厦门大学学报（自然科学版）》2002年第1期103-107,共5页Journal of Xiamen University：Natural Science

基　　金：福建省自然科学基金资助项目 (C982 0 0 0 5 )

摘　　要：应用RT RAPD技术 ,以 6bp随机引物反转录双孢蘑菇 (Agaricusbisporus) 0 2菌株常温培养和高温诱导菌丝体的总RNA ,10bp随机引物PCR扩增 ,得到几个高温诱导差异片段 ,对OPU13引物的差异片段 0 2U13克隆并测序 ,发现该片段可能编码tRNAVal(密码子GUC)基因 .推测GUC可能是稀有密码子 ,在某些耐温基因中出现频率较高 .识别该稀有密码子的tRNAVal在常温条件下不表达或表达量很少 ,而在高温诱导下引发一定的调控机制促使它们大量的合成 。Using the RT-RAPD method, total RNA was reverse-transcripted with 6 bp random primer, then cDNA was amplified by PCR with 10 bp random primer. Several related thermotolerance DNA fragments of strain 02 were obtained. The fragment 02U13 with OPU13 random primer was cloned and sequenced . It is found that 02U13 is probable a gene fragment of tRNA Val. GUC may be a rare codon, and it is abundant in some related themotolerance genes. The gene of tRNA Val for GUC is silence or low activity at normal temperature. But it express actively when the organism was induced at high temperature, and result of the high expression of the themotolerance related genes.

关 键 词：双孢蘑菇 RT-RAPD 耐温相关基因 tRNA^Val 食用菌 高温诱导 稀有密码子 调控机制 

分 类 号：Q756[生物学—分子生物学] S646.11[农业科学—蔬菜学]