Home > Browse Issues > Vol.35 No.2
Effect of SV40 Enhancer on Cow β-casein Promoter Activity
Zhong Yu1, Shao Zheng2, Jiang Liming3*
1Analysis Center of Guangdong Medical College, Zhanjiang 524023, China;
2Department of Parasitology, Guangdong Medical College, Zhanjiang 524023, China;
3Institute of Biochemistry and Molecular Biology, Guangdong Me
2Department of Parasitology, Guangdong Medical College, Zhanjiang 524023, China;
3Institute of Biochemistry and Molecular Biology, Guangdong Me
Abstract: By overlapping PCR method SV40 enhancer was inserted into the op0 promoter to construct recombinant
promoter op0-SV40enh， and its activity was analyzed. Firstly， PCR amplified 5′-2.1 Kb fragment of cow
β-casein promoter， 3′-1 Kb fragment of cow β-casein promoter and SV40 enhancer sequence. Secondly， three fragments
were connected by overlap PCR and inserted into multiply clone sites of pGL3-Basic vector and a recombinant
vector named pGL3-op0-SV40enh was constructed. Finally， the activity of op0 promoter and op0-SV40enh
promoter were detected by Dual-Luciferase Reporter Assay System. Overlapping PCR spliced out of the length of 3.4 Kb
fragments， and the sequencing results consistent with the expected results. These indicated that the recombinant
pGL3-op0-SV40enh vector be constructed successfully. The results of Dual-Luciferase Reporter Assay indicated
that Op0-SV40enh promoter activity well above op0 promoter activity. It could be conclusion that SV40 Enhancer
inserted into cow β-casein promoter op0 can significantly improve expression activity of luciferase report gene.
promoter op0-SV40enh， and its activity was analyzed. Firstly， PCR amplified 5′-2.1 Kb fragment of cow
β-casein promoter， 3′-1 Kb fragment of cow β-casein promoter and SV40 enhancer sequence. Secondly， three fragments
were connected by overlap PCR and inserted into multiply clone sites of pGL3-Basic vector and a recombinant
vector named pGL3-op0-SV40enh was constructed. Finally， the activity of op0 promoter and op0-SV40enh
promoter were detected by Dual-Luciferase Reporter Assay System. Overlapping PCR spliced out of the length of 3.4 Kb
fragments， and the sequencing results consistent with the expected results. These indicated that the recombinant
pGL3-op0-SV40enh vector be constructed successfully. The results of Dual-Luciferase Reporter Assay indicated
that Op0-SV40enh promoter activity well above op0 promoter activity. It could be conclusion that SV40 Enhancer
inserted into cow β-casein promoter op0 can significantly improve expression activity of luciferase report gene.
CN
EN