Construct synthetic gene encoding artificial spider dragline silk protein and its expression in milk of transgenic mice

被引:62
作者
Xu, Hong-Tao
Fan, Bao-Liang
Yu, Shu-Yang
Huang, Yin-Hua
Zhao, Zhi-Hui
Lian, Zheng-Xing
Dai, Yun-Ping
Wang, Li-Li
Liu, Zhao-Liang
Fei, Jing
Li, Ning [1 ]
机构
[1] China Agr Univ, Coll Biol Sci, State Key Labs Agrobiotechnol, Beijing 100094, Peoples R China
[2] Hebei Univ Engn, Handan, Hebei, Peoples R China
[3] China Agr Univ, State Key Lab Agrobiotechnol, Beijing, Peoples R China
[4] China Agr Univ, Coll Anim Sci & Technol, Beijing, Peoples R China
关键词
dragline silk; spider; transgenic mice;
D O I
10.1080/10495390601091024
中图分类号
S8 [畜牧、 动物医学、狩猎、蚕、蜂];
学科分类号
0905 ;
摘要
Based on the known partial cDNA sequence of dragline silk protein an artificial gene monomer, a 360 bp sequence, was designed and polymerized to encode an analog of dragline silk protein. Six tandem copies of monomer were cloned into pBC1 vector and microinjected into the pronuclei of fertilized Kunming White eggs. Transgenic mice were screened by Polymerase Chain Reaction (PCR) and Southern blot which revealed that 10 mice ( 5 male, 5 female) among 58 mice were transgenic positive. Milk of five F0 mice and eight F1 mice was analyzed by Western blot, and two F0 mice and seven F1 mice expressed recombinant dragline silk protein. In transgenic mice milk a maximum of concentration of recombinant dragline silk protein was 11.7 mg/ L by radioimmunoassay.
引用
收藏
页码:1 / 12
页数:12
相关论文
共 26 条
[1]   Purification and characterization of recombinant spider silk expressed in Escherichia coli [J].
Arcidiacono, S ;
Mello, C ;
Kaplan, D ;
Cheley, S ;
Bayley, H .
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 1998, 49 (01) :31-38
[2]   Synthetic spider dragline silk proteins and their production in Escherichia coli [J].
Fahnestock, SR ;
Irwin, SL .
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 1997, 47 (01) :23-32
[3]   Production of synthetic spider dragline silk protein in Pichia pastoris [J].
Fahnestock, SR ;
Bedzyk, LA .
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 1997, 47 (01) :33-39
[4]  
Foo CWP, 2002, ADV DRUG DELIVER REV, V54, P1131
[5]   Hypotheses that correlate the sequence, structure, and mechanical properties of spider silk proteins [J].
Hayashi, CY ;
Shipley, NH ;
Lewis, RV .
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES, 1999, 24 (2-3) :271-275
[6]   Evidence from flagelliform silk cDNA for the structural basis of elasticity and modular nature of spider silks [J].
Hayashi, CY ;
Lewis, RV .
JOURNAL OF MOLECULAR BIOLOGY, 1998, 275 (05) :773-784
[7]   Artificial fibrous proteins: A review [J].
Heslot, H .
BIOCHIMIE, 1998, 80 (01) :19-31
[8]   C-13 NMR of Nephila clavipes major ampullate silk gland [J].
Hijirida, DH ;
Do, KG ;
Michal, C ;
Wong, S ;
Zax, D ;
Jelinski, LW .
BIOPHYSICAL JOURNAL, 1996, 71 (06) :3442-3447
[9]  
Hinman M.B., 1992, J BIOL CHEM, V267, P19320
[10]   Synthetic spider silk: a modular fiber [J].
Hinman, MB ;
Jones, JA ;
Lewis, RV .
TRENDS IN BIOTECHNOLOGY, 2000, 18 (09) :374-379