Construction and Identification of a Yeast Two-Hybrid Bait Vector and Its Effect on the Growth of Yeast Cells and the Self-Activating Function of Reporter Genes for Screening of HPV18 E6-Interacting Protein

• Published in: Journal of Huazhong University of Science and Technology. Medical sciences Vol. 30; no. 1; pp. 8 - 12
• Main Author: 梅泉 李双 刘萍 奚玲 王世宣 孟玉菡 刘杰 杨欣慰 卢运萍 汪辉
• Format: Journal Article
• Language: English
• Published: Heidelberg Huazhong University of Science and Technology 01.02.2010; Department of Obstetrics and Gynecology,The Second People's Hospi%Cancer Biology Research Center,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China%Family Planning Research Institute,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China%Department of Obstetrics and Gynecology,The Second People's Hospital of Yichang City,Yichang 443000,China; Cancer Biology Research Center,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China
• Subjects: DNA-Binding Proteins - biosynthesis; DNA-Binding Proteins - genetics; Genes, Reporter - genetics; Genetic Vectors - genetics; HeLa Cells; Humans; Medicine; Medicine & Public Health; Oncogene Proteins, Viral - biosynthesis; Oncogene Proteins, Viral - genetics; Plasmids - genetics; Protein Interaction Mapping; Recombinant Proteins - biosynthesis; Recombinant Proteins - genetics; RNA, Messenger - biosynthesis; RNA, Messenger - genetics; Two-Hybrid System Techniques; Yeasts - genetics; Yeasts - growth & development; Yeasts - metabolism; HPV18 E6; yeast two-hybrid system; gene; bait plasmid
• ISSN: 1672-0733; 1993-1352
• DOI: 10.1007/s11596-010-0102-8

By using a yeast two-hybrid system,a yeast two-hybrid bait vector was constructed and identified for screening of the HPV18 E6-interacting proteins,and its effects on the growth of yeast cells and the activation of reporter genes were investigated.Total mRNA extracted from Hela cells was reversely transcribed into cDNA.Fragment of HPV18 E6 cDNA was amplified using RT-PCR and directly ligated to the pGBKT7 vector.The recombinant plasmid was confirmed by restriction endonuclease analysis and DNA sequencing.Th...