摘要
选用HTLV-Ⅲ前病毒核酸序列中的6333—6410作为合成HIV-env26肽结构基因,加上必要序列及接头共104bp,分成8个寡核苷酸片段,使结构基因能自身连接成多重串联体。用DNA合成仪合成。组装后与高效表达质粒pRC23重组,转化TAP106。经筛选、鉴定及DNA序列分析,得到一株含有HIV-env26肽基因4重串联体的重组菌,命名为pXY104。
A gene for HIV-env 26 peptide corresponding to the sequence from 6333 to 6410 of the provirus of HTLV-Ⅲ was synthesized. The gene was devided into eight fragments and sythesized with ABI 381A DNA sythesizer. With the operation of annealing and ligating, these fragments were assembled into a concatennated gene as a(b)n c (in this paper n = 4). Then the gene was inserted into plasmide pRC23 and the recombinants were used to transform E. coli TAP106.A recombinant colony has been obtained and identified by means of in situ hybridization, restriction enzyme mapping and DNA sequencing, showed that the gene of HIV-env 26 peptide has been inserted into the plasmid pRC23 as tetramer and the direction and reading frame of the inserted gene are correct. The recombinant plasmide was name pXY104.
