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人乙肝病毒增强子ⅡB1结合因子(hB1F)系FtzF1(NR5A)亚家族的新成员。经基因重组法将人hb1fcDNA置于小鼠白蛋白增强子/启动子序列下游构建成肝特异重组载体,通过原核显微注射将该载体导入小鼠受精卵原核,经注射且状态良好的卵回输至假孕母鼠输卵管。产下仔鼠经PCR和Southernblotting鉴定,同时RTPCR和Westernblotting分析转基因的表达。阳性Founder鼠与正常C57鼠交配以建立转基因纯系小鼠,F1代以PCR法鉴定。结果共获得4只PCR鉴定转基因阳性Founder鼠,其中一只同时经Southernblotting鉴定为阳性。RTPCR和Westernblotting结果显示,外源基因在转基因小鼠的肝组织成功表达。遗传学分析表明,转基因已整合入小鼠基因组并可稳定遗传。
A New Member of Human Fetal Hepatitis B Virus Enhancer IIB1 Binding Factor (hB1F) FtzF1 (NR5A) Subfamily. Recombinant human hb1f cDNA was placed downstream of the mouse albumin enhancer / promoter sequence by gene recombination to construct a liver-specific recombinant vector. The vector was introduced into the prokaryotic nucleus of the mouse by prokaryotic microinjection, Lost to the fake pregnant mouse fallopian tubes. The offspring were identified by PCR and Southern blotting, and the transgene expression was analyzed by RTPCR and Western blotting. Positive F immortalized mice were crossed with normal C57 mice to establish transgenic pure line mice. Fl generation was identified by PCR. Results A total of 4 PCR-positive transgenic Founder mice were obtained. One of them was positive by Southern blotting. The results of RTPCR and Western blotting showed that the foreign gene was successfully expressed in the liver tissue of transgenic mice. Genetic analysis showed that the transgene has been integrated into the mouse genome and can be stably inherited.