| 马小松王英振△ 王昌耀刘金钊.双基因真核表达载体pIRES-BMP2-TGFβ3 的构建与鉴定[J].,2011,11(9):1639-1642 |
| 双基因真核表达载体pIRES-BMP2-TGFβ3 的构建与鉴定 |
| Construction and Identification of PIRES-BMP2-TGFβ3 BicistronicEukayotic Expression Vector |
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| DOI： |
| 中文关键词: 骨形态发生蛋白 转化生长因子 双基因真核表达载体 |
| 英文关键词: BMP2 TGFβ3 Bicistronic eukaryotic expression vector |
| 基金项目: |
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| 中文摘要: |
| 目的：构建与鉴定骨形态发生蛋白BMP2 和转化生长因子TGFβ3 双基因真核表达载体pIRES-BMP2-TGFβ3。方法：首先，
用PCR 方法从质粒pGEMT/BMP2 中扩增出BMP2 基因全长，并将其连入双基因真核表达载体pIRES，得到质粒pIRES-BMP2，
其次，从人胚胎组织提取总RNA，反转录成cDNA，以反转录的cDNA 为模板，PCR 扩增出TGFβ3 基因全长，将TGFβ3 基因连
入质粒pIRES-BMP2；用酶切的方法筛选出阳性重组质粒，并进行测序鉴定。结果：酶切鉴定证明已将BMP2 和TGFβ3 两个基因
连入载体中，测序结果完全正确。结论：成功构建PIRES-BMP2/TGFβ3 双基因真核表达载体。 |
| 英文摘要: |
| Objective: To construct a bicistronic eukayotic expression vector pIRES-BMP2-TGF. Methods: The BMP2 gene was
obtained from pGEMT/BMP2 plasmid by PCR. And it was inserted into bicistronic eukaryotic expression plasmid vector pIRES. The
TGFβ3 was extracted from human embryonal tissue by RT-PCR, then the gene was inserted into the plasmid pIRES-BMP2. The inserted
target genes in the plasmid were detected by restriction enzyme digestion and nucleotide sequencing. Results: The direction and sequences
of the new bicistronic eukaryotic expression vector pIRES-BMP2-TGFβ3 were correct. Conclusion: The bicistronic eukaryotic
expression vector was successfully constructed. |
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