Expression and characterization of the hepatitis E virus ORF3 protein in the methylotrophic yeast, Pichia pastoris

被引：22

作者：

Lal, SK ^{[1
]}

Tulasiram, P ^{[1
]}

Jameel, S ^{[1
]}

机构：

[1] INT CTR GENET ENGN & BIOTECHNOL,VIROL GRP,NEW DELHI 110067,INDIA

来源：

GENE | 1997年 / 190卷 / 01期

关键词：

alcohol oxidase promoter; expression plasmid; integrative transformation; open reading frame; heterologous gene expression; methanol utilization pathway; glycosylation pattern; PUTATIVE STRUCTURAL PROTEINS; HIGH-LEVEL EXPRESSION; ANTIBODY-RESPONSES; GENE; CLONING; HEV;

D O I：

10.1016/S0378-1119(96)00698-1

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

We have used the methylotrophic yeast, Pichia pastoris, to express the open reading frame 3 (ORF3) of the hepatitis E virus (HEV). The ORF3 gene codes for a 123-amino-acid protein that contains highly immunodominant epitopes and is a potentially useful diagnostic and immunoprophylactic antigen. The expressed protein showed positive on immunoblots probed against antibodies raised in rabbit and infected human patient sera. In order to optimize the ORF3 protein expression, we have examined the regulated expression of this protein and characterized it. Unlike its expression in E. coli, the ORF3 protein was present in both the soluble and insoluble fractions of the cell lysate. The expressed protein is not glycosylated and does not undergo any major processing in the host strain.

引用

页码：63 / 67

页数：5

共 20 条

[1] EXPRESSION OF HEPATITIS-E VIRUS PUTATIVE STRUCTURAL PROTEINS IN RECOMBINANT VACCINIA VIRUSES [J].

CARL, M ;

ISAACS, SN ;

KAUR, M ;

KE, JK ;

TAM, AW ;

YARBOUGH, PO ;

REYES, GR .

CLINICAL AND DIAGNOSTIC LABORATORY IMMUNOLOGY, 1994, 1 (02) :253-256

[2] HIGH-LEVEL EXPRESSION OF TETANUS TOXIN FRAGMENT-C IN PICHIA-PASTORIS STRAINS CONTAINING MULTIPLE TANDEM INTEGRATIONS OF THE GENE [J].

CLARE, JJ ;

RAYMENT, FB ;

BALLANTINE, SP ;

SREEKRISHNA, K ;

ROMANOS, MA .

BIO-TECHNOLOGY, 1991, 9 (05) :455-460

[3] RECENT ADVANCES IN THE EXPRESSION OF FOREIGN GENES IN PICHIA-PASTORIS [J].

CREGG, JM ;

VEDVICK, TS ;

RASCHKE, WC .

BIO-TECHNOLOGY, 1993, 11 (08) :905-910

[4] HIGH-LEVEL EXPRESSION AND EFFICIENT ASSEMBLY OF HEPATITIS-B SURFACE-ANTIGEN IN THE METHYLOTROPHIC YEAST, PICHIA-PASTORIS [J].

CREGG, JM ;

TSCHOPP, JF ;

STILLMAN, C ;

SIEGEL, R ;

AKONG, M ;

CRAIG, WS ;

BUCKHOLZ, RG ;

MADDEN, KR ;

KELLARIS, PA ;

DAVIS, GR ;

SMILEY, BL ;

CRUZE, J ;

TORREGROSSA, R ;

VELICELEBI, G ;

THILL, GP .

BIO-TECHNOLOGY, 1987, 5 (05) :479-485

[5]

CREGG JM, 1987, BIOL RES IND YEASTS, V2, P1

[6] SOLID-PHASE ENZYME-LINKED-IMMUNOSORBENT-ASSAY FOR HEPATITIS-E VIRUS IGG AND IGM ANTIBODIES UTILIZING RECOMBINANT ANTIGENS AND SYNTHETIC PEPTIDES [J].

DAWSON, GJ ;

CHAU, KH ;

CABAL, CM ;

YARBOUGH, PO ;

REYES, GR ;

MUSHAHWAR, IK .

JOURNAL OF VIROLOGICAL METHODS, 1992, 38 (01) :175-186

[7]

Digane M.E., 1988, DEV IND MICROBIOL, P59

[8] ISOLATION OF ALCOHOL OXIDASE AND 2 OTHER METHANOL REGULATABLE GENES FROM THE YEAST PICHIA-PASTORIS [J].

ELLIS, SB ;

BRUST, PF ;

KOUTZ, PJ ;

WATERS, AF ;

HARPOLD, MM ;

GINGERAS, TR .

MOLECULAR AND CELLULAR BIOLOGY, 1985, 5 (05) :1111-1121

[9] EXPRESSION AND DIAGNOSTIC UTILITY OF HEPATITIS-E VIRUS PUTATIVE STRUCTURAL PROTEINS EXPRESSED IN INSECT CELLS [J].

HE, JK ;

TAM, AW ;

YARBOUGH, PO ;

REYES, GR ;

CARL, M .

JOURNAL OF CLINICAL MICROBIOLOGY, 1993, 31 (08) :2167-2173

[10] MOLECULAR-CLONING AND SEQUENCING OF THE MEXICO ISOLATE OF HEPATITIS-E VIRUS (HEV) [J].

HUANG, CC ;

NGUYEN, D ;

FERNANDEZ, J ;

YUN, KY ;

FRY, KE ;

BRADLEY, DW ;

TAM, AW ;

REYES, GR .

VIROLOGY, 1992, 191 (02) :550-558

← 1 2 →