A CONSTRUCTION AND AN ANALYSIS OF A GENE ENCODING SECRETORY LEUKOCYTE PROTEASE INHIBITOR AND PROTEIN DISULFIDE ISOMERASE USING A CO-EXPRESSION VECTOR IN SACCHAROMYCES CEREVISIAE BJ1824

Hermansyah, Hermansyah (2020) A CONSTRUCTION AND AN ANALYSIS OF A GENE ENCODING SECRETORY LEUKOCYTE PROTEASE INHIBITOR AND PROTEIN DISULFIDE ISOMERASE USING A CO-EXPRESSION VECTOR IN SACCHAROMYCES CEREVISIAE BJ1824. Journal of Chemical Technology and Metallurgy, 55 (6). pp. 1999-2008. ISSN 1314-7978

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Abstract

The secretory Leukocyte Protease Inhibitor (SLPI) is a protease inhibitor which can be found in saliva, bronchial mucus, seminal plasma and the amniotic membrane. It has been recognized for anti-inflammatory, antimicrobial and wound healing. Recombinant SLPI produced in Saccharomyces cerevisiae would be a useful biomaterial for wound healing. The yeast as a heterologous host holds an important role to express human SLPI. The yeast PDI is an enzyme which is required in the formation of disulfide bonds and folding of SLPI. A co-expression vector harboring SLPI gene and PDI1 gene is constructed aiming to strengthen the expression of SLPI in S. cerevisiae. The SLPI gene from the amniotic membrane is fused with HM1 signal peptide, namely hmSLPI. The HM1 signal peptide is used to mediate the translocation of SLPI across the membrane cells. The plasmids construction and characterization are done in Escherichia coli TOP10 prior to its transformation to S. cerevisiae BJ1824. The hmSLPI fusion gene and PDI1 gene are successfully amplified in sized 483 bp and 1,583 bp, respectively. The restriction analysis and nu�cleotide sequencing indicate that the recombinant plasmids, pAT_hmSLPI, pAT_PDI1 and pAT_PDI1_hmSLPI, are successfully constructed. All of the recombinant plasmids are also successfully introduced into S. cerevisiae BJ1824.

Item Type: Article
Subjects: Q Science > QD Chemistry > QD415-436 Biochemistry
Divisions: 08-Faculty of Mathematics and Natural Science > 47201-Chemistry (S1)
Depositing User: Hermansyah Hermansyah
Date Deposited: 20 Sep 2021 05:46
Last Modified: 20 Sep 2021 05:46
URI: http://repository.unsri.ac.id/id/eprint/53708

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