摘要
天花粉蛋白为单链核糖体失活蛋白,具糖苷酶活性,对 HIV 和多种动植物病毒有明显的抑制作用,有广泛的理论价值和应用前景.本文在国内首次采用聚合酶链式反应(PCR)方法,以栝楼染色体 DNA 为模板、合成的寡聚核苷酸为引物,扩增出了编码成熟天花粉蛋白的 DNA 片段(称 gene 1)和编码含23个氨基酸组成的分泌型信号肽、成熟天花粉蛋白以及羧基端延伸因子在内的 DNA 片段(称 gene 2).两 DNA
The genomic DNA was isolated and purified from fresh leaves of Trichosanthin Korilowii Maxim.With CTAB method.After digested with EcoRI,two forms gene were amplified by PCR with synthetic oligonucletides as primers.The first one contains a putative secretory signal peptide and a carboryl extension(named gene 1),and the second one is the gene alone (named gene 2).The two DNA fragments were cloned into vector pUC 18 and sequenced with the result that two genes have 99.30% homology comparated with the reported gene sequences.The expression of two genes in E.Celi was analysised with Western blotting and the results indicated that gene 2 can express some amount of fusion protein containing a part of α-peptide of lac Z gene and α-Trichosanthin,but gene 1 can't do so.
出处
《北京师范大学学报（自然科学版）》
CAS
CSCD
1992年第2期174-175,共2页
Journal of Beijing Normal University(Natural Science)
基金
国家自然科学基金
关键词
天花粉蛋白基因
克隆
序列
表达
a-Trichosanthin
gene
cloning
sequencing
expression
secretory signal peptide
