Screening method and application of recombinant pichia pastoris high-yield expression strain

• Main Authors: ZHANG JIANCHENG, CAO YUFENG, YI XIAONAN, SHI LI, CHEN XIAOYU
• Format: Patent
• Language: Chinese; English
• Published: 07.04.2023
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; COMPOSITIONS THEREOF; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; CULTURE MEDIA; ENZYMOLOGY; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; ORGANIC CHEMISTRY; PEPTIDES; PROCESSES OF PREPARING SUCH COMPOSITIONS; PROCESSES USING MICROORGANISMS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

The invention provides a screening method and application of a recombinant pichia pastoris high-yield expression strain, and relates to the technical field of bioengineering.The screening method comprises the steps that a fluorescent quantitative PCR method is adopted for obtaining GAPDH and AOX1 of recombinant pichia pastoris transformants subjected to induced expression and Ct values of target genes; the Mut < + > high-yield expression strain can be rapidly screened by comparing the size relationship of the Ct values of the GAPDH, the AOX1 and the target gene, and the technical problems of complex screening operation and low flux of the recombinant pichia pastoris high-yield expression strain in the prior art are relieved. 本发明提供了一种重组毕赤酵母高产表达株的筛选方法及其应用，涉及生物工程的技术领域，包括采用荧光定量PCR的方法获取经诱导表达的重组毕赤酵母转化子的GAPDH、AOX1和目的基因的Ct值，通过比较GAPDH、AOX1和目的基因的Ct值的大小关系可快速筛选获得Mut+高产表达株，缓解了现有技术中重组毕赤酵母高产表达株筛选操作复杂，通量低的技术问题。