【摘要】 取对虾(Penaeid shrimp)白斑综合征病毒(WSSV)基因组DNA随机文库中1个约8 kb的克隆片段进行测序。DNA序列用DNA star软件进行分析,共发现43个100bp以上阅读框(ORF),其中1条链31个,互补链12个。用BLAST软件将43个ORF及其推导的氨基酸序列分别与GenBank中核酸蛋白数据库序列进行相似性比较,结果无显著同源性。从白斑综合征病毒基因组的测序片段中,发现一个810bp阅读框,其编码氨基酸序列与海栖热袍菌(Thermotoga maritima)外膜蛋白具有24％的同源性,设计一对引物,扩增出该阅读框,用T载体克隆并按正确阅读框连接到E.coli表达载体pGEX-6p-1上。重组菌经IPTG诱导后,菌体SDS-PAGE显示有1条大小为56kD的融合表达蛋白产生。更多还原

【Abstract】 A genome DNA fragment about 8 kb from random library of white spot syndrome virus(WSSV)of shrimp (Penaeid shrimp) was sequenced and the DNA sequence was analyzed by DNA star software. Forty three open reading frames(ORF) over 100 bp in length were found, 31 ORFs in a chain, and 12 ORFs in the reverse complement chain. The ORFs and deduced amid acid sequence were compared to the database in GenBank by BLAST software respectively, and no obvious homology was found. From the genomic fragment of WSSV, an ORF of 810 bp in length which had 24% homology with outer membrane protein of Thermotoga maritima was amplified by PCR with specific primers. The PCR product was cloned with pUCm-T vector and then inserted into E.coli pGEX-6p-l vector with proper reading frame. SDS-PAGE of the recombinant BL21 bacteria showed that a fusion protein of 56 kD was expressed.更多还原