Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Method for microbe gene clone

A cloning method and microbial technology, applied in the field of genetics, can solve the problems of complicated workload, low exchange rate, strong subjectivity, etc., and achieve the effects of reducing risks, clear results, and broadening the scope of cloning and utilization.

Inactive Publication Date: 2013-01-30
JIANGHAN UNIVERSITY
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] (1) Microorganisms are small, so observation and identification are very difficult and highly subjective
However, linkage marker mapping requires identifying the phenotype of each strain in the segregating population, which is very complicated and difficult
[0006] (2) At present, the molecular markers developed for microorganisms are limited, which greatly restricts their application; in fine positioning, the available markers are less and less, or even no markers are available, making the work impossible. Even if there are markers, due to the low exchange rate , it is necessary to isolate and identify a large number of strains to obtain the genetic distance accurately, and the workload increases geometrically; only to find the segment containing the target gene, gene prediction is also required, and false positives or false negatives of gene prediction cannot be avoided

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for microbe gene clone
  • Method for microbe gene clone
  • Method for microbe gene clone

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] A method for microbial gene cloning, comprising the following steps (see figure 1 ):

[0025] (1) Construction of segregation population: use parent 1 and parent 2 with differences in target traits (such as toxicity, drug resistance, temperature sensitivity, etc.) to cross. If the microorganism is haploid, then F 1 It is a segregation group, if it is diploid, then go to F 2 Generations become a segregating population, which is the pre-screening population.

[0026] (2) Automatic screening of isolated populations: according to the characteristics of the target traits, select the selection pressure for the target traits (such as resistant plants, drugs, high temperature, etc.), automatically screen the isolated populations, and obtain the screened populations;

[0027] (3) DNA extraction: extract the genomic DNA of 2 parents, 2 mixed populations before and after screening;

[0028] (4) Determination of the target site: According to the high-throughput sequencing proce...

Embodiment 2

[0031] A kind of rice blast avirulent gene cloning method (see figure 2 ):

[0032](1) Identification of virulent and avirulent rice blast races. The physiological races of rice blast were isolated and cultured from the diseased leaves of Lijiang Xintuan Heigu rice in Enshi, Hubei Province, and the rice material IRBL 12 was inoculated with these physiological races. It was found that after inoculation with race S223, the typical rice blast fusiform appeared Lesions, indicating that the S223 race is virulent to IRBL 12. After inoculation with S12, IRBL 12 had almost no lesions, indicating that the S12 race was not toxic to IRBL 12.

[0033] (2)F 1 Obtaining and culturing hybrid ascospores. The parent strains of Magnaporthe grisea S223 and S12 were cultured on oatmeal medium (30g of oatmeal, 10g of sucrose, 18g of agar, 1000ml of water) at 25°C until the mycelium met, and continued to cultivate at 20°C under light until the junction of the mycelium appeared Ascus shell bla...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a method for microbe gene clone. The method comprises the steps of constructing segregation populations by hybridizing parents having target character differences; automatically screening the segregation populations by setting a selection pressure to obtain screened populations; high-throughput genome sequencing of the two parents, populations before screening and populations after screening; obtaining specific loci of the parents by preliminary direct assembly and comparison; comparing the specific loci of the parents with the genomes of the populations before screening and the populations after screening in a completely matching mode; looking for loci which exists in the parents and the populations before screening but disappears in the populations after screening, wherein the loci are candidate gene loci for controlling target characters; obtain full-length sequence of the genes by comparing with the genomes of the parents; cloning target genes via PCR amplification; and verifying functions of the cloned target genes via genetic complementary experiments. The method directly clones genes themselves, and reduces a lot of microspecies isolation and identification work of the microbes. The method has little workload, fast speed and low risk.

Description

technical field [0001] The invention belongs to the field of genetics and discloses a microbial gene cloning method. Background technique [0002] The types of microorganisms in nature are extremely complex, including bacteria, fungi, and viruses, and their hosts also include humans, animals, and plants. For humans, the functions of microbial genes are rich and diverse, both good and bad. For example, rhizobia and leguminous plants form root nodules symbiotically, and with the help of nitrogenase genes, nitrogen in the air can be fixed for plant nutrition, which can reduce the application of chemical fertilizers, protect environment. However, the rice blast caused by the virulence gene in Magnaporthe grisea is one of the most serious fungal diseases on rice, which can cause about 30% of rice yield reduction, or even no harvest, seriously threatening national food security. Therefore, cloning the toxicity genes and drug resistance genes of microorganisms is very important t...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12N15/10C12Q1/68
CPCC12Q1/6806C12Q1/68C12N15/10C12Q2527/156C12Q2535/125C12Q2539/101
Inventor 彭海张静章伟雄
Owner JIANGHAN UNIVERSITY
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com