CAJ | 학술논문

[目的]构建小鼠肌肉抑制素（MSTN）基因打靶载体。[方法]从小鼠后肢肌肉组织中提取总RNA并合成cDNA,以cDNA为模板克隆MSTN的编码区,以5’UTR及上游共3kb区域和3’UTR及下游共1.4kb区域分别作为长同源臂和短同源臂插入到质粒pBluescript_SK＋中,以neo基因和HSV-tk基因作为正负筛选基因构建MSTN基因的打靶载体pLoxP-5N3T-M。[结果]经酶切和测序鉴定,构建的小鼠MSTN基因打靶载体pLoxP-5N3T-M符合设计要求。[结论]成功构建小鼠MSTN基因打靶载体pLoxP-5N3T-M。
[목적]구건소서기육억제소（MSTN）기인타파재체。[방법]종소서후지기육조직중제취총RNA병합성cDNA,이cDNA위모판극륭MSTN적편마구,이5’UTR급상유공3kb구역화3’UTR급하유공1.4kb구역분별작위장동원비화단동원비삽입도질립pBluescript_SK＋중,이neo기인화HSV-tk기인작위정부사선기인구건MSTN기인적타파재체pLoxP-5N3T-M。[결과]경매절화측서감정,구건적소서MSTN기인타파재체pLoxP-5N3T-M부합설계요구。[결론]성공구건소서MSTN기인타파재체pLoxP-5N3T-M。
[Objective] This study aimed to construct Myostatin （MSTN） gene targeting vector of mouse. [Method] Total RNA was extracted from hindlimb muscle tissues of mouse to synthesize cDNA as the template to clone the coding region of MSTN. The CDS of MSTN gene including 3 kb 5’ homologous arm and 1.4 kb 3’ homologous arm were inserted into vector pBluescript_SK ＋ to construct the targeting vector pLoxP-5N3T-M. The neo and HSV-tk gene were cloned into vector pBluescript_SK＋ as positive and negative selective gene. [Result] Restriction enzyme digestion and sequencing results showed that mouse MSTN gene was cloned into the targeting vector pLoxP-5N3T-M. [Conclusion] The mouse MSTN gene targeting vector pLoxP5N3T-M was successfully constructed.