Reverse plant genetics is the most classical research idea in plant genetic engineering. It usually starts with a gene, regulates the expression level of one or more genes by means of one or more over-expression and interference expression, observes phenotype changes, and carries out a series of molecular mechanism research on related genes.

Construction of Overexpression Vector

The target gene fragments were synthesized by artificial gene synthesis or obtained directly from plant genome, and subcloned into plasmids with promoters, replicators, resistance markers and screening genes to achieve the overexpression of the target gene, so as to study the biological function of the target gene.

The main overexpression plasmid Zoonbio currently uses is pCAMBIA series, which contains one-element and two-element expression vectors. Based on the promoter of Cauliflower mosaic virus 35S (CaMV35S), the target gene is integrated into plants using T-DNA system based on Ti plasmid of Agrobacterium rhizobii, carrying Kana resistance and hygromycin screening markers on the vector. Genome, thus realizing the overexpression of target genes.

The vector picture information is as follows:


Construction of Interference Vector

In the study of gene function, the operation of over-expression of target genes is our frequent interference expression. In plant system, Zoonbio currently mainly uses pTCK303 series vectors based on Rice Intron to produce ShRNA (our company only undertakes scientific research projects, the ownership of plasmids and the corresponding commercial company).

The system also has the promoter of Cauliflower mosaic virus 35S (CaMV35S), carries Kana resistance and hygromycin screening markers on the vector, and also has GUS gene. We can construct complementary gene fragments according to the target gene sequence, and form shRNA structure in vivo, so as to achieve the inhibitory effect on the expression of the target gene.

The vector picture information is as follows:


Characteristic of Zoonbio Construction

1.According to the purpose of scientific research, provide professional construction design scheme. For example, the fusion site of GFP and the reservation of GUS gene are suggested.

2.In addition to delivering plasmid-containing E. coli, we can transform additional Agrobacterium tumefaciens such as EHA105 GV3101 Lba4404, and identify Agrobacterium tumefaciens by PCR to ensure the successful transformation of the target gene.

3.Only need to provide sequence, no need to provide other external materials, Zoonbio one-stop solution.


Related services:

suspension culture of plant cells service

model crop genetically modified service

subcellular localization of plant proteins service