Construct synthetic gene encoding artificial spider dragline silk protein and its expression in milk of transgenic mice.
ABSTRACT 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.
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ABSTRACT: Spider dragline silk is considered to be the toughest biopolymer on Earth due to an extraordinary combination of strength and elasticity. Moreover, silks are biocompatible and biodegradable protein-based materials. Recent advances in genetic engineering make it possible to produce recombinant silks in heterologous hosts, opening up opportunities for large-scale production of recombinant silks for various biomedical and material science applications. We review the current strategies to produce recombinant spider silks.Microbial Biotechnology 11/2013; 6(6):651-663. · 3.21 Impact Factor
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ABSTRACT: The potential of genetically modified organisms (GMOs) to increase allergenicity or predispositions to allergies has attracted much attention. An approach that will properly and holistically evaluate the allergenicity of GMOs is yet to be found. Here, 85 transgenes that have been reported in both international and domestic studies during recent years are summarized case by case; 49 of the transformed genes were from plant sources and 36 were from animal sources. EVALLER™, a web server for the in silico assessment of potential protein allergenicity, was used to evaluate the potential of the transgenic proteins as allergens. The biomedical journals listed in Highwire (http://highwire.stanford.edu/) were searched and reviewed to decipher whether any of the transformed genes were linked to allergenicity or human health. The EVALLER analysis identified 5 allergenic genes, whilst our literature review found 11 genes that were either related to allergic cases or to clinical adverse events; all 16 of these genes have been used in GMOs. The analysis pathway that we have developed can help guide the selection of genes to be used in genetic modification. The pathway also provides a paradigm for allergenicity analysis of transgene candidates.Chinese Science Bulletin 57(15). · 1.37 Impact Factor
- Acta Hydrobiologica Sinica 01/2010; 34(1):225-228.