摘要
目的构建针对大鼠结缔组织生长因子(connective tissue growth factor ,CTGF)的小分子干扰RNA(smal interfering RNA,siRNA)荧光逆转录病毒表达载体。方法将体外合成的两对各段含65碱基的寡核苷酸连接到pSIREN-RetroQ-ZsGreen载体,构建重组质粒,同时设立非特异对照。结果酶切、DNA序列鉴定均证实插入片段的正确性。结论成功地构建了鼠CTGF-siRNA荧光逆转录病毒表达载体。为进一步研究其对体内外抗纤维化作用打下基础。
Objective To design and construct retrovirus expressing smal interference RNAs(siRNAs)targeting rat us connective tissue growth factor(CTGF)mediated by pSIREN-RetroQ-ZsGreen vector.Methods Four pairs of 65 base-pair oligonucleotides were synthesized in vivo and inserted in pSIREN-RetroQ-ZsGreen vector,promoter to construct recombinant plasmids.Oligos with a scrambled sequence were used as a negat ive control.Results Correct construction of CTGF siRNAs vectors was identified by methods of restriction enzyme analysis,and nucleotide sequence determination.Conclusion The fluorescent retrovirus expression vectors of CTGF siRNAs were constructed successfully.This wil facilitate the stuy of antifibrotic intervention in vivo and in vitro.
作者
杨邱
刘伏友
彭佑铭
刘虹
肖力
YANG Qu;LIU Fu-you;PENG You-ming;LIU Hong;XIAO Li(Shenzhen City People's Hospital,Shenzhen 518000,Guangdong,China;The Second Xiangya Hospital,Central South University,Changsha 410002,Hunan,China)
