目的:构建鸡白细胞介素2(IL-2)真核重组表达质粒,体外鉴定其能否表达。方法:将鸡IL-2cDNA(RT-PCR法获得),克隆入真核高效表达质粒pcDNA3.1(+)中,构建pcDNA-IL-2重组表达质粒。PCR和双酶切的方法鉴定克隆的正确性。然后构建pcDNA IL GFP...
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目的:构建鸡白细胞介素2(IL-2)真核重组表达质粒,体外鉴定其能否表达。方法:将鸡IL-2cDNA(RT-PCR法获得),克隆入真核高效表达质粒pcDNA3.1(+)中,构建pcDNA-IL-2重组表达质粒。PCR和双酶切的方法鉴定克隆的正确性。然后构建pcDNA IL GFP重组表达质粒(即GFP基因与IL2的一段上游基因融合表达),PCR方法鉴定克隆的正确性。脂质体法转染COS1细胞,荧光显微镜下观察荧光。结果:正确构建了真核重组表达质粒pcDNA-IL2和pcDNA-IL-GFP。荧光显微镜下可观察到很强的绿色荧光。结论:鸡IL-2克隆和表达成功。为下一步用重组质粒pcDNA-IL2免疫BALB/c小鼠,研制鸡IL-2单克隆抗体奠定了基础。
Several H11N2 subtype of Avian influenza A viruses were isolated from aquatic birds in live bird markets when we surveyed the ecology of the influenza in East China for more than two years and identified by specific *...
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Several H11N2 subtype of Avian influenza A viruses were isolated from aquatic birds in live bird markets when we surveyed the ecology of the influenza in East China for more than two years and identified by specific *** hemagglutinin(HA) and neuraminidase(NA) genes of one representative virus named A/Duck/Yangzhou/44/2002(H11N2)(DYZ/44/02) was *** results showed the HA nucleotide sequence of DYZ/44/02 has high identity with Dk/England/56(H11N6) and the NA nucleotide sequence of DYZ/44/02 has more than 95% sequence homology with Dk/Hokkaido/49/98(H9N2).Sequencing and phylogenetic analysis of the N2 NA genes of DYZ/44/02 revealed that the NA gene of DYZ/44/02 has close relationships with that of Ck/Korea/MS96/96-like H9N2 virus and are distinct from those of Ck/Beijing/94(H9N2).The sequence of cleavage site of DYZ/44/02 consists of a single arginine,as is the case with most other hemagglutinins exhibiting low susceptibility to proteolytic activation.
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