Title:
RECOMBINANT VECTOR AND EXPRESSION PURIFICATION METHOD FOR HEAT-LABILE UNG FUSION PROTEIN
Document Type and Number:
WIPO Patent Application WO/2022/089573
Kind Code:
A1
Abstract:
Provided are a recombinant vector and an expression purification method for a heat-labile UNG fusion protein. The method comprises: cloning a gene sequence of Cod UNG to a pCold-sumo plasmid to construct a recombinant plasmid pCold-sumo-Cod UNG, transforming the recombinant plasmid into an Escherichia coli BL21 (DE3) competent cell, simultaneously transforming a plasmid pG-Tf2 which expresses chaperone into the competent cell, and carrying out low-temperature-induced expression to obtain a soluble sumo-Cod UNG fusion protein. According to the method, the problem of insoluble Cod UNG expression products is overcome.
Inventors:
ZHANG YONGYOU (CN)
WANG DAN (CN)
SONG NAJIE (CN)
WANG DAN (CN)
SONG NAJIE (CN)
Application Number:
PCT/CN2021/127313
Publication Date:
May 05, 2022
Filing Date:
October 29, 2021
Export Citation:
Assignee:
UNIV XIAMEN (CN)
XIAMEN ZEESAN BIOTECH CO LTD (CN)
XIAMEN ZEESAN BIOTECH CO LTD (CN)
International Classes:
C12N15/70; C12N9/24; C12N15/56
Foreign References:
| CN112342231A | 2021-02-09 | |||
| CN108504670A | 2018-09-07 | |||
| CN102250846A | 2011-11-23 | |||
| CN102839189A | 2012-12-26 |
Other References:
ANONYMOUS: "pColdS-SUMO", BIOFENG, BIOFENG, CN, 12 August 2017 (2017-08-12), CN, pages 1 - 2, XP055925804, Retrieved from the Internet [retrieved on 20220530]
DATABASE Nucleotide NCBI; 15 April 2005 (2005-04-15), ANONYMOUS : "Gadus morhua mRNA for uracil-DNA glycosylase precursor (ung1 gene)", XP055925808, Database accession no. AJ275971
ANDERSSON DANIEL, SVEC DAVID, PEDERSEN CATHRINE, HENRIKSEN JØRN, STÅHLBERG ANDERS: "Preamplification with dUTP and Cod UNG Enables Elimination of Contaminating Amplicons", INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, MOLECULAR DIVERSITY PRESERVATION INTERNATIONAL (MDPI), BASEL, CH, vol. 19, no. 10, 1 October 2018 (2018-10-01), Basel, CH , pages 3185, XP055925811, ISSN: 1661-6596, DOI: 10.3390/ijms19103185
See also references of EP 4239074A4
DATABASE Nucleotide NCBI; 15 April 2005 (2005-04-15), ANONYMOUS : "Gadus morhua mRNA for uracil-DNA glycosylase precursor (ung1 gene)", XP055925808, Database accession no. AJ275971
ANDERSSON DANIEL, SVEC DAVID, PEDERSEN CATHRINE, HENRIKSEN JØRN, STÅHLBERG ANDERS: "Preamplification with dUTP and Cod UNG Enables Elimination of Contaminating Amplicons", INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, MOLECULAR DIVERSITY PRESERVATION INTERNATIONAL (MDPI), BASEL, CH, vol. 19, no. 10, 1 October 2018 (2018-10-01), Basel, CH , pages 3185, XP055925811, ISSN: 1661-6596, DOI: 10.3390/ijms19103185
See also references of EP 4239074A4
Attorney, Agent or Firm:
SHOUCHUANG JUNHE PATENT AGENT CO., LTD. XIAMEN (CN)
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