4 results about "Gateway Technology" patented technology

This invention discloses a method for constructing a tomato core membrane-associated protein interaction library based on Gateway technology under Fusarium oxysporum infection conditions and its application, relating to the field of genetic engineering technology. The method includes: using tomato root tissue infected with Fusarium oxysporum as material, extracting total RNA and enriching mRNA; synthesizing first-strand cDNA using Biotin-attB2-Oligo(dT) primers; synthesizing double-stranded cDNA enzymatically; ligating it with a specific double-stranded attB1 adapter; introducing attB1 and attB2 sites at both ends of the cDNA; cloning the cDNA into the donor vector pDONR222 via BP recombination to construct a primary entry library; and recombinating the primary entry library with vectors pGADT7 and pPR3-N via LR recombination to construct a nuclear secondary library and a membrane secondary library, respectively. The constructed libraries have high titers, long insert fragments, and low empty vector rates. This invention also relates to the application of the library in screening the interaction between Fusarium oxysporum effector proteins and tomato nuclear membrane-associated proteins, providing an efficient and specific resource platform for studying plant-pathogen interaction mechanisms.

The application discloses a panax notoginseng phospholipase gene PnPLA1-21 and application thereof. The nucleotide sequence of the PnPLA1-21 gene is shown as SEQ ID NO:1, and the protein encoded by the PnPLA1-21 gene is shown as SEQ ID NO:2. The application clones the gene, constructs an RNAi carrier pHellsgate 2-PnPLA1-21 by using a Gateway technology, transforms the agrobacterium after the PnPLA1-21 gene in panax notoginseng leaves is instantaneously silenced, inoculates fusarium solani, and carries out resistance analysis. The results show that the disease spot area of the panax notoginseng leaves is significantly expanded after the RNAi silencing, is about 2 times of that of a control group, and the expression level of the PnPLA1-21 is positively correlated with the disease resistance. The application proves for the first time that the PnPLA1-21 positively regulates the resistance of panax notoginseng to the root rot pathogenic bacteria, and can be used for cultivating transgenic panax notoginseng plants resistant to the root rot, and provides new gene resources and technical support for panax notoginseng disease resistance breeding.

The invention relates to the technical field of molecular biology, in particular to an arabidopsis thaliana circadian rhythm expression system, a construction method and application, and the system comprises a carrier I and a carrier II; the first vector is composed of a pCPE-Dest vector skeleton, an AtCAT3 promoter of an arabidopsis thaliana endogenesis and a target gene and can drive high expression of the gene at night. And the vector II is composed of a pCPE-Dest vector skeleton, an AtCAB2 promoter of an endogenous arabidopsis thaliana and a target gene, and can drive the daytime high expression of the gene. The system allows the to-be-detected gene to be expressed in arabidopsis thaliana in different day and night expression time phases under the driving of different promoters, and can directly explore the influence on plant physiological activities when the gene expression rhythm is changed; a to-be-detected gene is allowed to be replaced at multiple cloning sites through an enzyme digestion ligation reaction, and promoters with different expression time phases are replaced by using a Gateway technology to drive gene expression. The experiment is simple and the cost is low.

The application provides application of UGT5 genes and SCPL12 genes in production of agrostemma herb base in hairy roots. The application is realized by designing primer compositions to amplify the UGT5 genes and the SCPL12 genes, constructing a recombination carrier by using a Gateway technology, transforming the recombination carrier into Agrobacterium rhizogenes to obtain an Agrobacterium rhizogenes strain containing the UGT5 and the SCPL12, infecting leaves of Leonurus japonicus by using the Agrobacterium rhizogenes strain, culturing to obtain transgenic Leonurus japonicus hairy roots, and further producing the agrostemma herb base in the hairy roots. The content of the agrostemma herb base in the hairy roots of Leonurus japonicus obtained by using the Agrobacterium rhizogenes mediation and simultaneously overexpressing the UGT5 and the SCPL12 is higher than that of a control group. The method lays a foundation for production of the agrostemma herb base in transgenic plant cells.