Dual-luciferase reporter gene vector for insect cells, construction method, recombinant vector and application

Abstract

The invention belongs to the field of reporter gene vectors, and particularly relates to a dual-luciferase reporter gene vector for insect cells, a construction method, a recombinant vector and application. The dual luciferase reporter gene vector for the insect cells is obtained by modifying a psiCHECK-2 vector; the transformation comprises the following steps: replacing an original SV40 promoter with an OpIE2 promoter, and inserting a synthesized DNA (Deoxyribose Nucleic Acid) fragment between an HSV-TK promoter and a Synthetic poly (A); the downstream of the hRluc gene of the psiCHECK-2 vector is used for inserting an insect target gene of interest; the nucleotide sequence of the synthesized DNA fragment is as shown in SEQ ID NO: 1. The dual-luciferase reporter gene vector for insect cells provided by the invention ensures powerful gene transcription and mRNA processing in insect cells, and is suitable for insect gene function research.

Description

technical field [0001] The invention belongs to the field of reporter gene carriers, and in particular relates to a dual-luciferase reporter gene carrier for insect cells, a construction method, a recombinant carrier and applications. Background technique [0002] Insects are the most numerous organisms in the world, with a large number and strong adaptability to the environment. Because of the advantages of easy access to materials, simple genetic manipulation, and obvious genetic phenotypes, it is favored by researchers and plays an important role in many aspects such as biomedicine, insecticides, and basic research. With the continuous development of molecular biology techniques, more than 800 insect cell lines have been established, which are often used in the research of immunofluorescence, protein interaction, and regulation of target genes by small molecule RNA. [0003] MiRNAs are non-coding RNA molecules of about 22 nucleotides (nt), which can degrade target mRNA o...

AI Technical Summary

Problems solved by technology

The co-transfection experiment has the following defects: First, the length of the full-length 3'-UTR of most target genes varies from several hundred to several thousand bases, and the recombinant reporter vector is constructed

[0006] In the prior art, although there is a Psicheck-2 vector design, which can monitor changes in gene expression by fusing firefly and sea cucumber luciferase reporter genes, this vector is almost non-existent in insect cells due to the species-specificity of the promoter. Activity, can not normally start the expression of luciferase gene on the carrier, so the use in insect cells is limited

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