| 1 | A novel aberrant splicing of G_(sa) transcript in human leukemia cell lines显示文摘The α subunit of the stimulatory G protein, Gsα, is involved in the stimulation of adenylate cyclase pathway of signal transduction. The various Gsα mRNAs are generated either by alternative splicing or by using alternative promoter. A novel aberrant splicing of Gsα transcript in human leukemia cell lines is reported. The entire coding region of Gsα gene obtained, referred to as GsαLeu, was amplified by PCR with cDNA from Jurkat human leukemia cDNA library or the reverse-transcribed first strand cDNA from human leukemia cell line HL-60 as a template. The result of DNA sequencing indicated that,compared with originally reported Gsα, GsαLeu has two in-frame deletions, one in amino acid residues 23—249 and the other in amino acid residues 254—260, suggesting that G(sαLeu) may play an important role in leukemia. The complete coding region of GsαLu was inserted downstream of the thrombin site of pGEX-2T fusion protein expression vector and expressed in the form of GST/G( | YE Qinong, YAO Xiao, WANG Hengliang, ZHANG Shu, LIU Huaitian, SU Guofu, HUANG Cuifen and ZHOU Tingchong Beijing Institute of Biotechnology, Beijing 100071, China Department of Biology, Shaanxi Normal College, Xi’an 710062, China Beijing Medical University, School of Oncology, Beijing 100036, China Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China Beijing Institute of Basic Medicine Sciences , Beijing 100850, China | 1999 | Chinese Science Bulletin1999，44，4： | 0 |
| 2 | Preliminary study on discovery of a novel molecule that interacts with CPP32 using two-hybrid system显示文摘Of ICE protease family, CPP32 (apopain or Yama) plays a central role in different apoptotic pathways. To study the molecules regulating CPP32, yeast two_hybrid system was used to identify proteins (peptides) that interact with CPP32. First, the CPP32 gene was cloned into plasmid vector pGBT9. The resulting recombinant plasmid was designated as pGBT9/CPP. The pGBT9 /CPP plasmid was transformed into the yeast strain HF7C, then the leukemia library was introduced. The transformation mixture was plated on medium lacking Trp, Leu and His in the initial screen. Colonies growing on the selection medium were further assayed for β galactosidase activity. Within 42 Trp +Leu +His + colonies only 5 turned blue in the presence of X_Gal. Plasmid DNA from 5 positive yeast colonies was prepared respectively and used to transform HB101 by electroporation. The transformation mixture was plated on medium lacking Leu to be selected for the library plasmid. Finally, only one library plasmid, designated as pY1, was determined to be truly positive by retransformation of pGBT9/CPP and the library plasmid into HF7C. The inserted cDNA of pY1 encodes a peptide of 15 amino acids, suggesting that it may be the domain interacting with CPP32. | Ye Qinong Yao Xiao Wang Hengliang Su Guofu Huang Cuifen Zhou Tingchong | 1998 | Chinese Science Bulletin1998，43，4： | 0 |
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