23 Jul Construction method of luciferase plasmid containing bovine fatty acid transport protein 1 gene promoter
Abstract
The invention discloses a construction method of a luciferase plasmid containing bovine fatty acid transport protein 1 gene promoter. The method comprises the following steps: obtaining the sequence of a 5′ flanking promoter of bovine FATP 1 gene first exon by a molecular cloning method, subcloning to a pMD19-T-Promoter carrier, then carrying out double-enzyme digestion reactions by utilizing KpnI and XhoI restriction enzymes, purifying and recycling the reaction product, grafting the product onto pGL3-Basic plasmid, converting escherichia coli JM109, selecting the identified positive clones, and carrying out a large amount of preparation and preservation. The luciferase plasmid (pGL3-Basic-Promoter), which is constructed by the method and contains a 5′ flanking promoter of bovine FATP 1 gene first exon, can transfect cells and carry out promoter functional activity analysis to select high-efficient tissue specific promoters, and overcomes the defects of tissue specific promoters shortage and weak regulation and control ability in the present animal gene engineering.
Year: 2014
Country: CN
Doc No: 103509811