摘要
为了研究梅花鹿S100A4(S100 calcium binding protein A4)基因在鹿茸生长过程中的作用。用RT-PCR法从生茸骨膜细胞总RNA中克隆了梅花鹿S100A4基因,在NCBI中对基因序列进行比对;将完整的基因序列与逆转录病毒表达载体pLEGFP-C1重组,获得了重组质粒pLEGFP-S100;用脂质体法将pLEGFP-S100与pVSV-G(被膜载体)共转染包装细胞GP2-293,获得重组病毒上清液,感染角柄骨膜细胞后逆转录病毒携带的基因进入宿主细胞。结果显示:S100A4基因是一个相对保守的基因,与多个物种的匹配度达到90%;重组逆转录病毒载体pLEGFP-S100可以形成重组逆转录病毒粒子,将S100A4基因导入靶细胞,并表达S100A4与GFP(Green fluores-cent protein)的融合蛋白。
In order to better understand the function of S100 calcium binding protein A4 in antler development of sika deer（Cervus nippon）,we cloned S100A4 genes from total RNA of cultured antlerogenic periosteum cells using reverse transcription polymerase chain reaction（RT-PCR）,S100A4 gene sequences were compared with closely related animal species in NCBI.Full lengths of S100A4 genes were inserted into a vector plasmid pLEGFP-C1（retroviral express vector）.The recombinant plasmid pLEGFP-S100 and pVSV-G（envelope vector） were co-transfected into GP2-293 cells（packaging cell line） using lipofectimin 2000,and the resultant viral supernatants were collected.The cultured pedicle periosteum cells were then infected with virus in the supernatants.Results showed that S100A4 gene was a relatively conserved gene,and had about 90% homology with several species.Recombinant retroviral vector pLEGFP-S100 could effectively deliver a gene into target cell line,and express a fusion GFP（green fluorescent protein）protein.
出处
《兽类学报》
CAS
CSCD
北大核心
2011年第1期103-107,共5页
Acta Theriologica Sinica
基金
国家自然科学基金资助项目(309771664)
