摘要
目的分离小鼠ING4(inhibitorofgrowthfamily,member4)基因并构建真核表达载体pcDNA3.1(+)ING4。方法从小鼠肝组织中提取总RNA,通过RTPCR扩增出ING4cDNA,构建真核表达载体pcDNA3.1(+)ING4,分别用双酶切、PCR、基因测序进行鉴定。结果RTPCR产物为约750bp的条带,构建的真核表达载体pcDNA3.1(+)ING4经双酶切、PCR鉴定均出现750bp大小的条带,基因测序正确。结论分离得到了小鼠ING4基因并成功构建真核表达载体pcDNA3.1(+)ING4。
Objective To isolate the gene encoding mouse ING4 and construct pcDNA3.1(+)-ING4 recombiant eukaryotic expression plasmid.Methods Using RT-PCR method the cDNA encoding the mouse ING4 was isolated with total RNA extracted from mouse liver tissue.The cDNA fragment was subcloned into the eukaryotic expression vector pcDNA3.1(+).Analysis by PCR、restricting enzyme digestion and DNA sequencing were carried out to demonstrate the sequence of the plasmid.Results RT-PCR product is about 750bp specific fragment;analysis was conducted by restricting enzyme digestion and PCR of pcDNA3.1(+)-ING4 recombiant plasmid showed that results were about 750bp. DNA sequencing revealed that ING4 cloning was successful.Conclusion The gene encoding mouse ING4 and construction of pcDNA3.1(+)- ING4 eukaryotic expression vector are successfully obtained.
出处
《苏州大学学报（医学版）》
CAS
北大核心
2005年第3期409-411,415,共4页
Suzhou University Journal of Medical Science
