Construction of a reporter gene vector for the full-length sequence of axonal motor protein Kif1a promoter gene
- VernacularTitle: 神经轴突运动蛋白Kif1a启动子基因全长序列报告基因载体构建
- Author:
Wei JIANG
1
;
Qing REN
1
;
Guanshuyuan LAI
1
Author Information
- Publication Type:Journal Article
- Keywords: kif1a ； Luciferase expression vector
- From: Journal of Apoplexy and Nervous Diseases 2021;38(4):292-295
- CountryChina
- Language:Chinese
- Abstract: Objective To clone full length of Kif1a gene promoter,and to construct and identify Kif1a gene promoter pGL3-Kif1a vectors.Methods Recombinant of pGEM-T easy vector (2565 bp) including Kif1a gene promoter full sequence was used as a template in the polymerase chain reaction (PCR) to amplify its promoter sequence (1853 bp).The PCR product was directly cloned into the luciferase reporter vector pGL3-Basic.Then the products were identified by DNA sequencing,nest PCR and restriction enzyme digestion.Then it was transfected into SCG cells,and the luciferase activity of the cells was analyzed after transfection.Results Restriction enzyme digestion and DNA sequencing confirmed that the sequenced segment (1853 bp) in the recombinant was identical to that in GenBank and the segment was inserted in right direction.The activity of pGL3-Kif1a was significantly higher than that in pGL3-basic vectors.Conclusion The luciferase expression vector-pGL3-Kif1a containing Kif1a gene promoter full length sequence is constructed successfully.The construction of pGL3-Kif1a recombinant plasmid lay a foundation of the analysis of promoter activities,gene expression regulatory mechanism and signal transduction of Kif1a.
- Full text:2024073012570442030Construction of a reporter gene vector for the full-length sequence of axonal motor protein Kif1a promoter gene.pdf
