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[Construction and Identification of the Bait Vector Containing Duck Circovirus Cap Gene for the Yeast Two-hybrid System]. 2015

Yu Xu, and Zhilong Zhang, and Yanyan Lu, and Lei Zhang, and Pengfei Li, and Renyong Jia

To construct a bait expression vector containing the duck circovirus Cap gene for use in the yeast two-hybrid system, the whole cap codon-optimized gene was inserted into pGBKT7 vector and confirmed by PCR, restriction enzyme digestion, and sequence analysis. After transformation into a Y2HGold yeast strain, the expression of Cap protein was analyzed by Western blotting. Toxicity and self-activation of the bait protein were detected using different dropout minimal base. PCR reaction, restriction enzyme digestion, and sequencing analyses indicated that the duck circovirus Cap gene was correctly inserted into pG- BKT7. Western blotting showed that the whole Cap protein was expressed. The recombinant bait protein had no toxicity and self-activation. Therefore, the bait vector with the Cap gene was constructed successfully, providing a foundation for future screening for interacting proteins in the yeast two-hybrid system.

UI MeSH Term Description Entries
D011993 Recombinant Fusion Proteins Recombinant proteins produced by the GENETIC TRANSLATION of fused genes formed by the combination of NUCLEIC ACID REGULATORY SEQUENCES of one or more genes with the protein coding sequences of one or more genes. Fusion Proteins, Recombinant,Recombinant Chimeric Protein,Recombinant Fusion Protein,Recombinant Hybrid Protein,Chimeric Proteins, Recombinant,Hybrid Proteins, Recombinant,Recombinant Chimeric Proteins,Recombinant Hybrid Proteins,Chimeric Protein, Recombinant,Fusion Protein, Recombinant,Hybrid Protein, Recombinant,Protein, Recombinant Chimeric,Protein, Recombinant Fusion,Protein, Recombinant Hybrid,Proteins, Recombinant Chimeric,Proteins, Recombinant Fusion,Proteins, Recombinant Hybrid
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
D004372 Ducks A water bird in the order Anseriformes (subfamily Anatinae (true ducks)) with a broad blunt bill, short legs, webbed feet, and a waddling gait. Duck
D005822 Genetic Vectors DNA molecules capable of autonomous replication within a host cell and into which other DNA sequences can be inserted and thus amplified. Many are derived from PLASMIDS; BACTERIOPHAGES; or VIRUSES. They are used for transporting foreign genes into recipient cells. Genetic vectors possess a functional replicator site and contain GENETIC MARKERS to facilitate their selective recognition. Cloning Vectors,Shuttle Vectors,Vectors, Genetic,Cloning Vector,Genetic Vector,Shuttle Vector,Vector, Cloning,Vector, Genetic,Vector, Shuttle,Vectors, Cloning,Vectors, Shuttle
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D012441 Saccharomyces cerevisiae A species of the genus SACCHAROMYCES, family Saccharomycetaceae, order Saccharomycetales, known as "baker's" or "brewer's" yeast. The dried form is used as a dietary supplement. Baker's Yeast,Brewer's Yeast,Candida robusta,S. cerevisiae,Saccharomyces capensis,Saccharomyces italicus,Saccharomyces oviformis,Saccharomyces uvarum var. melibiosus,Yeast, Baker's,Yeast, Brewer's,Baker Yeast,S cerevisiae,Baker's Yeasts,Yeast, Baker
D017925 Circovirus A genus of the family CIRCOVIRIDAE that infects SWINE; PSITTACINES; and non-psittacine BIRDS. Species include Beak and feather disease virus causing a fatal disease in psittacine birds, and Porcine circovirus causing postweaning multisystemic wasting syndrome in pigs (PORCINE POSTWEANING MULTISYSTEMIC WASTING SYNDROME). Beak and feather disease virus,Circovirus, Porcine,Porcine Circovirus,Circoviruses, Porcine,Porcine Circoviruses
D020798 Two-Hybrid System Techniques Screening techniques first developed in yeast to identify genes encoding interacting proteins. Variations are used to evaluate interplay between proteins and other molecules. Two-hybrid techniques refer to analysis for protein-protein interactions, one-hybrid for DNA-protein interactions, three-hybrid interactions for RNA-protein interactions or ligand-based interactions. Reverse n-hybrid techniques refer to analysis for mutations or other small molecules that dissociate known interactions. One-Hybrid System Techniques,Reverse One-Hybrid System Techniques,Reverse Two-Hybrid System Techniques,Three-Hybrid System Techniques,Yeast Two-Hybrid Assay,Yeast Two-Hybrid System Techniques,One-Hybrid System Technics,Reverse Three-Hybrid System Techniques,Three-Hybrid System Technics,Tri-Hybrid System Techniques,Two-Hybrid Assay,Two-Hybrid Method,Two-Hybrid System Technics,Yeast One-Hybrid System Techniques,Yeast Three-Hybrid Assay,Yeast Three-Hybrid System,Yeast Three-Hybrid System Techniques,Yeast Two-Hybrid System,n-Hybrid System Techniques,Assay, Two-Hybrid,Assay, Yeast Three-Hybrid,Assay, Yeast Two-Hybrid,Assays, Two-Hybrid,Assays, Yeast Three-Hybrid,Assays, Yeast Two-Hybrid,Method, Two-Hybrid,Methods, Two-Hybrid,One Hybrid System Technics,One Hybrid System Techniques,One-Hybrid System Technic,One-Hybrid System Technique,Reverse One Hybrid System Techniques,Reverse Three Hybrid System Techniques,Reverse Two Hybrid System Techniques,System Technique, n-Hybrid,System Techniques, n-Hybrid,System, Yeast Three-Hybrid,System, Yeast Two-Hybrid,Systems, Yeast Three-Hybrid,Systems, Yeast Two-Hybrid,Technic, One-Hybrid System,Technic, Three-Hybrid System,Technic, Two-Hybrid System,Technics, One-Hybrid System,Technics, Three-Hybrid System,Technics, Two-Hybrid System,Technique, One-Hybrid System,Technique, Three-Hybrid System,Technique, Tri-Hybrid System,Technique, Two-Hybrid System,Technique, n-Hybrid System,Techniques, One-Hybrid System,Techniques, Three-Hybrid System,Techniques, Tri-Hybrid System,Techniques, Two-Hybrid System,Techniques, n-Hybrid System,Three Hybrid System Technics,Three Hybrid System Techniques,Three-Hybrid Assay, Yeast,Three-Hybrid Assays, Yeast,Three-Hybrid System Technic,Three-Hybrid System Technique,Three-Hybrid System, Yeast,Three-Hybrid Systems, Yeast,Tri Hybrid System Techniques,Tri-Hybrid System Technique,Two Hybrid Assay,Two Hybrid Method,Two Hybrid System Technics,Two Hybrid System Techniques,Two-Hybrid Assay, Yeast,Two-Hybrid Assays,Two-Hybrid Assays, Yeast,Two-Hybrid Methods,Two-Hybrid System Technic,Two-Hybrid System Technique,Two-Hybrid System, Yeast,Two-Hybrid Systems, Yeast,Yeast One Hybrid System Techniques,Yeast Three Hybrid Assay,Yeast Three Hybrid System,Yeast Three Hybrid System Techniques,Yeast Three-Hybrid Assays,Yeast Three-Hybrid Systems,Yeast Two Hybrid Assay,Yeast Two Hybrid System,Yeast Two Hybrid System Techniques,Yeast Two-Hybrid Assays,Yeast Two-Hybrid Systems,n Hybrid System Techniques,n-Hybrid System Technique
D036022 Capsid Proteins Proteins that form the CAPSID of VIRUSES. Procapsid Protein,Procapsid Proteins,Viral Coat Protein,Viral Coat Proteins,Viral V Antigens,Viral V Proteins,Capsid Protein,Viral Outer Coat Protein,Antigens, Viral V,Coat Protein, Viral,V Antigens, Viral,V Proteins, Viral

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