[A preliminary study of the structure prediction and expression of SARS-CoV spike protein].
In this study, the encoding sequences of SARS-CoV spike protein were analyzed by bioinformatics methods, the structural characteristics and functions were forecasted based on available data. It suggests that the fragment of spike glycoprotein (S401-659) may be crucial for viral attachment and may be a major immunodominant epitope. Then the fragment was amplified and subcloned into expression vector pET28a(+) and pPIC9K. These two plasmids pET28a(+)-S and pPIC9K-S were transformed to E.coli strain BL21(DE3)-star and Pichia pastoris, respectively. SDS-PAGE and Western blot analysis showed that the recombinant protein was successfully expressed. The denatured inclusion bodies were purified with Ni-NTA chelate agarose and its purity can reach 90%.
Sun
Mei-Ling
ML
College of Animal Science and Technology , Northwest Sci-Tech University of Agriculture and Forestry, Yangling, Shaanxi, 712100, China. lisasun78@yahoo.com.cn
Piao
Jin-Hua
JH
Yin
Chang-Cheng
CC
Yang
Gong-She
GS
Huang
Hua-Liang
HL
D015153:Blotting, Western; D000939:Epitopes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D015967:Gene Expression Regulation, Viral; D005822:Genetic Vectors / Q000235:genetics; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010843:Pichia / Q000235:genetics; D021382:Protein Sorting Signals / Q000235:genetics; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism*; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics / Q000378:metabolism*; D064370:Spike Glycoprotein, Coronavirus; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D000939:Epitopes; D008562:Membrane Glycoproteins; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D064370:Spike Glycoprotein, Coronavirus; D014759:Viral Envelope Proteins