摘要
以分离自马铃薯主栽品种“紫花白”的马铃薯卷叶病毒(PLRV)分离物的RNA为模板,用人工合成的引物,用反转录和随后PCR扩增的方法合成了PLRV外壳蛋白(CP)基因的cDNA,并克隆于pUC19中。进一步用限制酶切和核苷酸序列分析表明,合成的cDNA由627个核苷酸组成(包括起始和终止密码),序列中有Hinc Ⅱ和BamH Ⅰ两个酶切位点,与国外报道一致。和国外的4个PLRV分离物CP基因序列对比结果,具有高度同源性,其同源率达99.0—99.7％。
A coat protein gene eDNA of a PLRV isolate infected commercial potato cultivar 'Purple Flower White' was synthesized by reverse transcription followed by Polymerase Chain Reaction amplification with synthesized 20merand 30mer primers. The synthesized eDNA was cloned in plasmid pUC19 in JM103. The full length CP gene eDNA clones pLCP2 and pLCP4 were further identified by restriction mapping analysis and, the nucleotide sequence of cDNA cloned in pLCP2 was analyzed with ABI 370 DNA sequence Tests show that the eDNA of this Chinese PLRV isolate CP gene consists of 627 nueleotides including start and stop codons with two restriction sites, HineⅡ and BamH I. Sequence analysis revealed that there is a high homology in nucleotide sequence in comparision with other PLRV isolates reported by Kawehuk et al., (1989), Prill et al., (1989), Mayo et al., (1989) and Smith et al., (1990).
出处
《中国病毒学》
CAS
CSCD
1992年第4期432-435,共4页
Virologica Sinica
基金
国家自然科学基金
关键词
马铃薯
卷叶病毒
外壳蛋白基因
Potato leaf roll virus(PLRV) Coat protein gene Molecular cloning Sequence analysis PCR
