A signal sequence trap based on cell enrichment using Anti-CD19 antibody coated magnetic beads

被引:3
作者
Rian, E [1 ]
Wabakken, T [1 ]
Hauge, H [1 ]
Aasheim, HC [1 ]
机构
[1] Norwegian Radium Hosp, Dept Immunol, N-0310 Oslo, Norway
关键词
D O I
10.1046/j.1365-3083.2001.00925.x
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
Precursors of many secreted and cell surface proteins contain NH2-terminal signal sequences that lead proteins into the endoplasmic reticulum and to the cell surface. Methods have been developed to clone and identify such proteins by trapping their NH2-terminal signal sequences, so called signal sequence traps. In this study we present an alternative and simplified signal sequence trap based on the combination of a novel vector construct expressing a cDNA library in fusion with a CD19 reporter gene, transfection in mammalian cells and selection of cells expressing trapped signal sequences using magnetic beads. Using this method we have isolated several known and novel factors with signal peptides.
引用
收藏
页码:280 / 284
页数:5
相关论文
共 15 条
[1]   Gapped BLAST and PSI-BLAST: a new generation of protein database search programs [J].
Altschul, SF ;
Madden, TL ;
Schaffer, AA ;
Zhang, JH ;
Zhang, Z ;
Miller, W ;
Lipman, DJ .
NUCLEIC ACIDS RESEARCH, 1997, 25 (17) :3389-3402
[2]   A new signal sequence trap using alkaline phosphatase as a reporter [J].
Chen, HC ;
Leder, P .
NUCLEIC ACIDS RESEARCH, 1999, 27 (04) :1219-1222
[4]   Molecular cloning of a novel T cell-directed CC chemokine expressed in thymus by signal sequence trap using Epstein-Barr virus vector [J].
Imai, T ;
Yoshida, T ;
Baba, M ;
Nishimura, M ;
Kakizaki, M ;
Yoshie, O .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1996, 271 (35) :21514-21521
[5]   Selection for genes encoding secreted proteins and receptors [J].
Klein, RD ;
Gu, QM ;
Goddard, A ;
Rosenthal, A .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1996, 93 (14) :7108-7113
[6]   A signal sequence trap based on a constitutively active cytokine receptor [J].
Kojima, T ;
Kitamura, T .
NATURE BIOTECHNOLOGY, 1999, 17 (05) :487-490
[7]   A SIMPLE METHOD FOR DISPLAYING THE HYDROPATHIC CHARACTER OF A PROTEIN [J].
KYTE, J ;
DOOLITTLE, RF .
JOURNAL OF MOLECULAR BIOLOGY, 1982, 157 (01) :105-132
[8]   Isolation and characterization of CA XIV, a novel membrane-bound carbonic anhydrase from mouse kidney [J].
Mori, K ;
Ogawa, Y ;
Ebihara, K ;
Tamura, N ;
Tashiro, K ;
Kuwahara, T ;
Mukoyama, M ;
Sugawara, A ;
Ozaki, S ;
Tanaka, I ;
Nakao, K .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1999, 274 (22) :15701-15705
[9]  
OKABE M, 1987, BLOOD, V69, P990
[10]   Identification of stromal cell products that interact with pre-B cells [J].
Oritani, K ;
Kincade, PW .
JOURNAL OF CELL BIOLOGY, 1996, 134 (03) :771-782