几种PCR方法在狂犬病毒基因测序中的应用.pdfVIP

2006, 22 ( 3) Chinese J urnal f Z n ses 225 : 1002- 2694( 2006) 03- 0225- 03 几种PCR 方法在狂犬病毒基因测序中的应用* 1 1 2 1 郭 利, 赵云蛟 , 李泽涛 , 钱爱东 : 本研究在狂犬病毒全基因测序中应用了RTPCR 法递减 PCR 法嵌套PCR 方法及 3Full RA CE 等方 法, 有效的解决了狂犬病毒在体 扩增时遇到的困难成功地获得了狂犬病毒野毒株BRV 的全基因的cDNA , 为实现基因测 序及下一步研究工作奠定了理论技术基础 : 狂犬病毒; PCR ; 基因测序 : R3739 : A Application of several PCR methods in the in vitro amplification of the rabies virus gene sequences GUO Li, ZHAO Yunjia , LI Zeta , QIAN Aid ng ( Colleg e of A n imal Science and Tech nology , Jilin A g ricult ural Un iv er sity , Jilin Changchun 130 118, China) ABSTRACT:In rder t res lve the pr blems enc untered in the in v itr o amplificati n f the rabies virus gene sequences, several PCR meth ds including RTPCR , decreased PCR , nested PCR and 3Full RA CE w ere applied f r study . It was f und that the w ild t ype rabies virus BRV strain c uld be btained , thus pr viding the technical basis f r further studies n the rabies virus genes. KE WORDS:rabies virus; PCR; survey c mplete sequence , cDNA 1. 2 PCR , , 1. 2. 1 RTPCR : RNA , Triz l RNA , ( 3g ) , Triz l , RNA - 20 RTPCR cDNA RNA , mRNA, , P1: 5 - GCACGACAGTT CCT CACCCTT CT- , 3 : 7000~ 7023 PCR P2: 5- GACCCAT T CT GAACAT CCTACCT T A- 3 : 8909~ 8932 PCR , PCR L , 1 932 bp, 1 cDNA , 42 60 min 1. 1 BRV : PCR : 94 40s56 60s72 3 min ( ) 30 JM 109 1. 2. 2 PCR , RNA T riz lTaq Plus Rnasin