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

Micrococcus glutamine synthetase gene as well as encoding protein and cloning method thereof

A glutamine and synthase technology, applied in the field of glutamine synthase gene, can solve the problems of low nitrogen fertilizer utilization rate, water environment pollution and the like

Inactive Publication Date: 2014-08-06
SHANGHAI UNIV
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, at present, the low utilization rate of nitrogen fertilizer and the problem of water pollution caused by excessive application have become a worldwide problem.

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
  • Micrococcus glutamine synthetase gene as well as encoding protein and cloning method thereof
  • Micrococcus glutamine synthetase gene as well as encoding protein and cloning method thereof
  • Micrococcus glutamine synthetase gene as well as encoding protein and cloning method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment one: Mico3 Cloning and analysis of the full-length coding region of the gene:

[0022] Micrococcus ( Micrococcus spp.) are mainly distributed in soil and water. Collect a small amount of river bank soil samples, take 1 gram of soil, and extract its metagenomic DNA. By analyzing the glutamine synthetase gene sequence of Micrococcus bacteria in the NCBI database, the primers were designed: Micoc1F: 5'-ATGTTCACCTCCCCCGCGGA-3' and Micoc1R: 5'-TCAGACCGAGTAGTAGAGCT-3'. Using this pair of primers, a glutamine synthetase gene was amplified by PCR from the extracted soil microbial metagenomic DNA, named as Mico3 . The gene was inserted into the cloning vector pMD18-T, the corresponding positive clone was selected, and the full-length DNA sequence of the gene was carried out to obtain the DNA sequence containing the complete ORF of the gene. The sequencing results showed that: Mico3 The full length of the ORF is 1425bp. The analysis result of DNAstar softwar...

Embodiment 2

[0023] Embodiment two: Mico3 Functional analysis in E. coli mutants

[0024] Through DNA sequencing analysis, select Mico3 Gene ORF transcription direction and lactose promoter on vector pMD18-T lacZ The clones with the same transcription direction were extracted and transformed into Escherichia coli mutant ET6017 (Genotype: F-,[araD139]B / r,Δ(argF-lac)169,flhD5301,Δ(fruK-yeiR) 725 (fruA25), relA1, rpsL150 (strR), Δ (glnG-glnA) 229, ha-10, deoC1). See literature: Merida, A., Flores E., Florencio F.J., Regulation of Anabaena sp. strain PCC7120 glutamine synthetase activity in a Synechocystis sp. strain PCC6803 derivative strain bearing the Anabaena glnA gene and a mutated host glnA gene. J Bacteri 1992, 174(2): 650-654. Due to the deletion of the gene encoding glutamine synthetase, the mutant cannot grow normally in the medium without glutamine. This strain was purchased from E. coli Genetic Stock Center, Yale University. Depend on Figure 4 and Figure 5 It shows tha...

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

PropertyMeasurementUnit
molecular weightaaaaaaaaaa
Login to View More

Abstract

The utility model relates to a micrococcus glutamine synthetase gene as well as encoding protein and a cloning method thereof. The gene comes from micrococcus bacteria and is a base sequence represented by SEQIDNO1. The gene encodes glutamine synthetase and can remarkably improve the capacity for synthesizing glutamine by model organism escherichia coli. Accordingly, the disclosed full-length gene and amino acid sequence have application values in the genetic engineering aspects of increasing the nitrogen utilization rate and the nitrogen content of an organism.

Description

technical field [0001] The invention relates to a glutamine synthetase gene, its encoded protein and its cloning method. In particular, a micrococcal glutamine synthetase gene, its encoded protein and its cloning method. Background technique [0002] Nitrogen is the mineral element most needed by organisms, and it is used to synthesize amino acids, nucleotides and various nitrogen-containing compounds required by organisms. Nitrogen utilization plays a very important role in crop production, and plant nitrogen metabolism and its regulation have attracted worldwide attention. However, at present, the low utilization rate of nitrogen fertilizer and the water pollution caused by excessive application have become a worldwide problem. Therefore, it is now necessary to reduce the amount of nitrogen fertilizer used, and to find plant strains with higher nitrogen utilization efficiency, so as to reduce the amount of nitrogen fertilizer used without reducing the yield. [0003] Th...

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
Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/52C12N9/00C12N15/70C12N1/21C12N15/10C12R1/19
Inventor 朱晨光王菁张贵敏凌慧玲唐远平梅冰宋任涛
Owner SHANGHAI UNIV
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