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Screening interactive proteins TEB4 by yeast two-hybride system

Lei Xu

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Abstract

【Objective】 To screen the interactive proteins of TEB4(MARCH-VI)and construct the vector.【Methods】Yeast two-hybrid technique was performed to screen a human lymphoma cDNA library with pGBKT7TEB4 as bait plasmid.About one hundred and fifty positive clones were obtained from SD/-4 medium plate.The plasmids from yeast were transformed into E.coli DH5α and extracted for sequencing.After repetitive hybrid confirmation,the putative protein that can interact with TEB4 was ATP synthase F0 subunit 6.This gene was amplified by PCR method and cloned into the pCMV-HA vector,harvesting a recombinant plasmid pCMV-HA-ATP-C6.【Results】Only one positive clone identified by yeast two-hybrid,a recombinant plasmid pCMV-HA-ATP-C6 was successful constructed.【Conclusion】Screen and initially identify the interactive protein of TEB4 is a foundation for the study on the protein of TEB4.

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What this paper is about

【Objective】 To screen the interactive proteins of TEB4(MARCH-VI)and construct the vector.【Methods】Yeast two-hybrid technique was performed to screen a human lymphoma cDNA library with pGBKT7TEB4 as bait plasmid.About one hundred and fifty positive clones were obtained from SD/-4 medium plate.The plasmids from yeast were transformed into E.coli DH5α and extracted for sequencing.After repetitive hybrid confirmation,the putative protein that can interact with TEB4 was ATP synthase F0 subunit 6.This gene was amplified by PCR method and cloned into the pCMV-HA vector,harvesting a recombinant plasmid pCMV-HA-ATP-C6.【Results】Only one positive clone identified by yeast two-hybrid,a recombinant plasmid pCMV-HA-ATP-C6 was successful constructed.【Conclusion】Screen and initially identify the interactive protein of TEB4 is a foundation for the study on the protein of TEB4.

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Available abstract

【Objective】 To screen the interactive proteins of TEB4(MARCH-VI)and construct the vector.【Methods】Yeast two-hybrid technique was performed to screen a human lymphoma cDNA library with pGBKT7TEB4 as bait plasmid.About one hundred and fifty positive clones were obtained from SD/-4 medium plate.The plasmids from yeast were transformed into E.coli DH5α and extracted for sequencing.After repetitive hybrid confirmation,the putative protein that can interact with TEB4 was ATP synthase F0 subunit 6.This gene was amplified by PCR method and cloned into the pCMV-HA vector,harvesting a recombinant plasmid pCMV-HA-ATP-C6.【Results】Only one positive clone identified by yeast two-hybrid,a recombinant plasmid pCMV-HA-ATP-C6 was successful constructed.【Conclusion】Screen and initially identify the interactive protein of TEB4 is a foundation for the study on the protein of TEB4.

Key concepts: Plasmid, Recombinant DNA, Yeast, clone (Java method), Biology, Two-hybrid screening, cDNA library, Molecular biology

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