Biomaterial Database

[Expression and preliminary applications of hepatitis B surface antigen mutants]. 2007

Chang-Cheng Zhu, and Chun-Zhen Luo, and Man Wang

Biochemistry and Molecular Biology, East China University of Science and Technology, Shanghai 200237, China. changchengzh@163.com

OBJECTIVE To clone and express HBsAg mutant in the Pichia pastoris. METHODS The cloned wild type pGAP-S was used as the DNA template to generate mutant type pGAP-MS with a single or double nucleotide changes incorporated in complementary oligonucleotide primers. The product was linearized with BspH I and transformed into Pichia pastoris strain GS115, and stable multicopy integrants were screened in medium containing different concentrations of Zeocin. RESULTS The pGAP-MS expression vector was successfully constructed and stable numbers integrated strains with high copy number were obtained. The expression of HBsAg mutant protein was identified by SDS-PAGE and Western blot with specific polyclonal antibody. The molecular weight of recombinant HBsAg mutant was 38 kDa. AxSYM HBsAg V2(Abbott)assays demonstrated all 10 HBsAg mutants were reactive. CONCLUSIONS The recombinant HBsAg mutant with immunoreactivity was successfully expressed in Pichia pastoris, and it was of practical value on the quality control and clinical applications of commercial assays.

UI	MeSH Term	Description	Entries
D009154	Mutation	Any detectable and heritable change in the genetic material that causes a change in the GENOTYPE and which is transmitted to daughter cells and to succeeding generations.	Mutations
D010843	Pichia	Yeast-like ascomycetous fungi of the family Saccharomycetaceae, order SACCHAROMYCETALES isolated from exuded tree sap.	Hansenula,Hansenulas,Pichias
D011994	Recombinant Proteins	Proteins prepared by recombinant DNA technology.	Biosynthetic Protein,Biosynthetic Proteins,DNA Recombinant Proteins,Recombinant Protein,Proteins, Biosynthetic,Proteins, Recombinant DNA,DNA Proteins, Recombinant,Protein, Biosynthetic,Protein, Recombinant,Proteins, DNA Recombinant,Proteins, Recombinant,Recombinant DNA Proteins,Recombinant Proteins, DNA
D003001	Cloning, Molecular	The insertion of recombinant DNA molecules from prokaryotic and/or eukaryotic sources into a replicating vehicle, such as a plasmid or virus vector, and the introduction of the resultant hybrid molecules into recipient cells without altering the viability of those cells.	Molecular Cloning
D006511	Hepatitis B Antigens	Antigens of the virion of the HEPATITIS B VIRUS or the Dane particle, its surface (HEPATITIS B SURFACE ANTIGENS), core (HEPATITIS B CORE ANTIGENS), and other associated antigens, including the HEPATITIS B E ANTIGENS.	HBAg,Hepatitis B Antigen,Antigen, Hepatitis B,Antigens, Hepatitis B,B Antigen, Hepatitis,B Antigens, Hepatitis
D000954	Antigens, Surface	Antigens on surfaces of cells, including infectious or foreign cells or viruses. They are usually protein-containing groups on cell membranes or walls and may be isolated.	Cell Surface Antigens,Surface Antigens,Surface Markers, Immunological,Cell Surface Antigen,Immunologic Surface Markers,Markers, Immunological Surface,Surface Antigen,Surface Markers, Immunologic,Antigen, Cell Surface,Antigen, Surface,Antigens, Cell Surface,Immunological Surface Markers,Markers, Immunologic Surface,Surface Antigen, Cell,Surface Antigens, Cell
D013997	Time Factors	Elements of limited time intervals, contributing to particular results or situations.	Time Series,Factor, Time,Time Factor
D015870	Gene Expression	The phenotypic manifestation of a gene or genes by the processes of GENETIC TRANSCRIPTION and GENETIC TRANSLATION.	Expression, Gene,Expressions, Gene,Gene Expressions
D017422	Sequence Analysis, DNA	A multistage process that includes cloning, physical mapping, subcloning, determination of the DNA SEQUENCE, and information analysis.	DNA Sequence Analysis,Sequence Determination, DNA,Analysis, DNA Sequence,DNA Sequence Determination,DNA Sequence Determinations,DNA Sequencing,Determination, DNA Sequence,Determinations, DNA Sequence,Sequence Determinations, DNA,Analyses, DNA Sequence,DNA Sequence Analyses,Sequence Analyses, DNA,Sequencing, DNA
D050505	Mutant Proteins	Proteins produced from GENES that have acquired MUTATIONS.	Mutant Protein,Protein, Mutant,Proteins, Mutant