本研究參照GenBank中編碼禽源熱休克蛋白HSP70、HSP40 和核上習糖體蛋白RL4等的基因序列，分别設計特異性引物，采用RT-PCR擴增HSP7開我0、HSP40和RPL4基因，經(jīng)雙酶切後(hòu)克隆到木費真核表達載體pEF1α-Myc，得到重組質粒pEF1α-Myc我要-HSP70、pEF1α-Myc-HSP40和pEF1α-到房Myc-RPL4；將(jiāng)構建好(hǎo)的睡森重組質粒，轉染HEK293T細胞後(hòu)，采用間接藍愛免疫熒光和Western-blot技術，對(duì)目的蛋白進(jì購裡n)行驗證。結果表明，Myc-HSP70、Myc-HSP40和M睡票yc-RPL4融合蛋白在HEK293T細胞用問内得到了正确表達。本研究爲後(hò購路u)續禽源HSP70、HSP40和RPL4基因的吃暗功能(néng)研究奠定了基礎。

Cloning and Expression of Avian-o跳算rigin Genes of HSP70，HSP40 and RPL4舞煙

In this study，specific primers were 件下respectively designed according to 年市the gene sequences of th會票e heat-shock protein（HSP70 a間去nd HSP40）and the 可金ribosomal protein（RPL討服4）registered in GenBank，then上輛 the genes of HSP70，HSP40 and能藍 RPL4 were amplifie不多d by RT-PCR，and after double e了友nzyme digestion，the哥國 qualified genes were cloned i錯答nto the eukaryotic expressi門白on plasmid，pEF1α-Myc，to obtain the re河輛combinant plasmids including p現低EF1α-Myc-HSP70，pEF1α-Myc-HSP40 and pEF1動身α-Myc-RPL4；after the那爸 recombinant plasmids were 微動transfected into HEK293T cell跳笑s，the expressed protein was ve快拿rified by indirect immunoflu光用orescence and Western-b身地lot. The results showed that th農黃e recombinant proteins of Myc-HSP國公70，Myc-HSP40 and Myc-RPL4 were corr姐姐ectly expressed船問 in HEK293T cells. Therefore，the 物用foundation was laid for fu如遠rther study on th件妹e genes of HSP70，HSP40 and RPL4.

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