Construct synthetic gene encoding artificial spider dragline silk protein and its expression in milk of transgenic mice.
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.
Xu
Hong-Tao
HT
State Key Laboratory for Agrobiotechnology, China Agriculture University, Beijing, PR China.
Fan
Bao-Liang
BL
Yu
Shu-Yang
SY
Huang
Yin-Hua
YH
Zhao
Zhi-Hui
ZH
Lian
Zheng-Xing
ZX
Dai
Yun-Ping
YP
Wang
Li-Li
LL
Liu
Zhao-Liang
ZL
Fei
Jing
J
Li
Ning
N
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern / Q000662:veterinary; D015153:Blotting, Western / Q000662:veterinary; D003001:Cloning, Molecular / Q000379:methods; D004247:DNA / Q000737:chemistry / Q000235:genetics; D005260:Female; D005349:Fibroins / Q000096:biosynthesis / Q000235:genetics*; D005813:Genes, Synthetic; D005822:Genetic Vectors / Q000235:genetics; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D008892:Milk / Q000378:metabolism / Q000502:physiology*; D008969:Molecular Sequence Data; D011247:Pregnancy; D011863:Radioimmunoassay; D013112:Spiders / Q000235:genetics*
C076815:spidroin 2; D004247:DNA; D005349:Fibroins