A kind of avermectin producing bacteria and its preparation method and application
A kind of abamectin, the technology of producing bacteria, applied in the biological field, can solve the problems such as complex processing process, low yield and the like
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0139] Construction of Abamectin Biosynthesis Gene aveD Inactivation (aveD Point Mutation) Strain
[0140] 1. Construction of recombinant plasmids for aveD point mutation
[0141] The primer sequences of the upstream DNA fragments of the aveD gene in the avermectin biosynthetic gene cluster were cloned by PCR as follows:
[0142] Primer 1: 5'-TAT GAA TTC CCT CGT CGA GGT GGC CGA G-3' (the underline is the EcoRI site) (SEQ ID NO: 4)
[0143] Primer 2: 5'-TTA AAG CTT ACC GCA GCC GAC GTC CAG G-3' (underlined is HindIII) (SEQ ID NO: 5)
[0144] The primer sequences of the downstream DNA fragments of the aveD gene in the avermectin biosynthetic gene cluster cloned by PCR are as follows:
[0145] Primer 3: 5'-TTA AAG CTT AAG CCG GCG GTG CGG CTC G-3' (underlined as HindIII) (SEQ ID NO: 6)
[0146] Primer 4: 5'-TTA TCT AGA TCC TCA CCC TTT CCC CCG GC-3' (underline is XbaI) (SEQID NO: 7)
[0147]Using the extracted total DNA of the avermectin-producing bacteria Streptomyc...
Embodiment 2
[0158] Construction of aveC aveD double mutant strain of abamectin biosynthesis gene
[0159] 1. Construction of aveC in-frame knockout plasmid
[0160] The primer sequence of the upstream DNA fragment of the aveC gene in the avermectin biosynthetic gene cluster cloned by PCR is as follows:
[0161] Primer 7: 5'-TTA GAA TTC CAT CAC GCT GCT GGA CTT CG-3' (the underline is the EcoRI site) (SEQ ID NO: 10)
[0162] Primer 8: 5'-ACG C CT GCA G GG CCA GAA ACA G-3' (underlined is PstI) (SEQ ID NO: 11)
[0163] The primer sequences of the downstream DNA fragments of the aveC gene in the avemectin biosynthetic gene cluster were cloned by PCR as follows:
[0164] Primer 9: 5'-TAT CTG CAG GTG AAT GCC GTG ATG TTC CTC-3' (underline is PstI) (SEQID NO: 12)
[0165] Primer 10: 5'-TAT TCT AGA TGC TCT CCT CCA CCA CAT CCT C-3' (XbaI is underlined) (SEQ ID NO: 13)
[0166] Using the extracted total DNA of the avermectin-producing bacteria StreptomycesavermectiniusATCC31267 as a te...
Embodiment 3
[0175] Construction of meiC heterologous anaplerotic strain
[0176] 1. Construction of plasmids for heterologous complementation of meiC
[0177] The primer sequences for PCR cloning of meilingmycin biosynthetic gene meiC are as follows:
[0178] Primer 13: 5'-TAT GGA TCC CTG CGC AAT AGG CTC ACC AC-3' (the underline is the BamHI site) (SEQ ID NO: 16)
[0179] Primer 14: 5'-TAT TCT AGA CGG GAG GTG TCT ACA AGT GC-3' (the underline is the XbaI site) (SEQ ID NO: 17)
[0180] Using the extracted total DNA of the meilingmycin-producing bacterium Streptomyces nanchangensis as a template, the meilingmycin biosynthetic gene meiC was amplified by PCR with the above-mentioned pair of specific primers, and a 1.7kb complete meiC gene was obtained Fragmented PCR products. After the PCR product was digested with BamH-XbaI, it was connected to the commercially available BamH-XbaI of pSET152 (an integrated E. coli-Streptomyces shuttle plasmid, see US1,171,583) containing erythromycin...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
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