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[马铃薯卷叶病毒中国分离物ORF2a基因的结构特征]

[Structure characteristics of ORF2a gene of potato leafroll virus Chinese isolate].

作者信息

Zhao Guo-Fen, Zhang He-Ling, Hasi Agula

机构信息

Department of Biology, Inner Mongolia University, Hohhot 010021, China.

出版信息

Sheng Wu Gong Cheng Xue Bao. 2002 Nov;18(6):744-8.

Abstract

According to the genomic sequence of foreign four PLRV isolates, three pairs of specific primer were designed and synthesized. The cDNA of the ORF2a gene of PLRV-Ch was synthesized by reverse transcription and followed by Polymerase Chain Reaction amplication. The synthesized 3' and 5' cDNA fragment of the PLRV-Ch ORF2a gene were inserted into pUC19 and cloned in E. coli JM109 and were sequenced respectively. The middle cDNA fragment were directly sequenced. The homology of nucleotide sequence of PLRV-Ch compared with PLRV-S (Scotland, UK), PLRV-N(Netherlands), PLRV-A(Australia) and PLRV-C(Canada) were 98.96%, 98.70%, 94.79%, 97.5%, the homology of putative amino acid sequence are 97.97%, 97.97%, 89.69%, 95.94%. In 3' region of ORF2a gene a slippery sequence for-1 frameshift and its downstream "stem-loop" or "pseudoknot" and upstream nucleotide sequence repeats were found. Authors suggested that the nucleotide repeat sequences characteristic for PLRV could form a tight successively folded complementary double stranded regions and hairpins. This structure possibly has something to do with-1 frameshift. The amino acid sequence of C terminus region of 70 kD protein translated by motif IV has a protease characteristic motif and a helicase motif IV. The amino acid sequence of polypeptide translated by ORF2a gene undergoing frameshift has a single-stranded nucleic acid binding protein-like characteristic motif.

摘要

根据国外4个马铃薯卷叶病毒(PLRV)分离株的基因组序列,设计并合成了3对特异性引物。通过反转录合成PLRV-Ch的ORF2a基因的cDNA,随后进行聚合酶链反应扩增。将合成的PLRV-Ch ORF2a基因的3'和5' cDNA片段插入pUC19并克隆到大肠杆菌JM109中,分别进行测序。中间的cDNA片段直接测序。PLRV-Ch与PLRV-S(英国苏格兰)、PLRV-N(荷兰)、PLRV-A(澳大利亚)和PLRV-C(加拿大)的核苷酸序列同源性分别为98.96%、98.70%、94.79%、97.5%,推测的氨基酸序列同源性分别为97.97%、97.97%、89.69%、95.94%。在ORF2a基因的3'区域发现了一个用于-1移码的滑序列及其下游的“茎环”或“假结”以及上游核苷酸序列重复。作者认为,PLRV特有的核苷酸重复序列可形成紧密连续折叠的互补双链区域和发夹结构。这种结构可能与-1移码有关。由基序IV翻译的70 kD蛋白C末端区域的氨基酸序列具有蛋白酶特征基序和解旋酶基序IV。由发生移码的ORF2a基因翻译的多肽的氨基酸序列具有单链核酸结合蛋白样特征基序。

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