马铃薯纺锤形块茎类病毒基因的化学合成与克隆

CHEMICAL SYNTHESIS AND CLONING OF A GENE FOR POTATO SPINDLE TUBER VIROID

马铃薯纺锤形块茎类病毒(Potato spindle tuber viroid,PSTV)是一种由359个核苷酸组成的植物类病毒。本文介绍用化学法合成PSTV基因的工作。将整个基因分成173、168和18bp 3个片段,对两个大片段分别进行克隆,最后得到含有PSTY基因的单链DNA。 首先,根据PSTV的核苷酸序列得到其cDNA,将整个cDNA分成A、B、C3段,分别为1-173、174-341和342-359。为了下一步将cDNA转录到RNA的研究,在每一个片段的3’

potato spindle tuber viroid ( PSTV ) is a plant viroid. It contains 359 nucleotides. The whole of gene was divided into three parts. Those parts contain the respective structural gene of 173bp, 163bp and l8bp.The gene fragment was chemically synthesized by DNA synthesizer. The synthetic gene was inserted into plasmid M13mp18, and transformed into JM109 strain. The clones containing the anticipated gene sequence was obtained by hybridization. The accuracy of sequence was confirmed by DNA sequencing.