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Objective To construct the recombinant plasmid pYES6-S and express and purify spike protein of severe acute respiratory syndrome(SANS)coronavirus in Saccharomyces cerevisiae. Methods DNA fragments of SANS coronavirus were obtained by reverse transeription.Four over- lapped fragments of spike protein genes were amplified by polymerase chain reaction(PCR)and ligated into an integral spike protein gene by restriction enzyme digestion.The spike protein gene recombined with pYES6 and cloned into E.coll.The recombinant plasmid pYES6-S was induced and expressed in Saccharomyces cerevisiae(INVScl)by galactose.Results The recombinant plasmid pYES6-S was confirmed that inserted fragment was right in length,direction and base matching by restriction enzyme digestion and sequencing.The purified protein encoded by the whole spike protein gene was about Mr 110?10~3 identified by electrophoresis.Conclusion The whole spike protein gene of SARS coronavirus is cloned into E.coli and the protein is expressed in Saccharomyces cerevisiae successful ly.which can be helpful in SARS vaccine research.
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Objective In order to find the polymorphism site applicable to efficient genetic diagnosis on Haemophilia A in Han Chinese Population in Wenzhou.Methods With the method of polymerase chain reaction(PCR) and polyacrylamide gel electrophoresis(PAGE),288 of X chromosomes from 96 men and 96 women were detected on the polymorphism of BCL I in the intron 18 of FV Ⅲ gene.Results The gene frequency of the polymorphic site BCL I was 34.38% in Han Chinese population in Wenzhou.43.75% women were heterozygous and the polymorphism information content(PIC) was 0.4512.Conclusion For Han Chinese population in Wenzhou,the BCL I genetic site has enough information,being one of the genetic markers with high polymorphism,applicable to the screening for carrier and prenatal diagnosis of Haemophilia A in Wenzhou.
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OBJECTIVE@#To study genetic polymorphism of D7S2846, D19S400 and D18S535 in Han population in Wenzhou.@*METHODS@#DNA was extracted with chelex-100 method from EDTA-blood samples of 194 unrelated individuals in Wenzhou and amplified with PCR technique. The PCR products were analyzed by PAG vertical electrophoresis and silver-stain.@*RESULTS@#6 alleles and 15 genotypes of D7S2846, 10 alleles and 36 genotypes of D19S400, 8 alleles and 26 genotypes of D18S535 was observed. The heterozygosities of the three loci are 0.644, 0.724 and 0.772; The power of discriminating are 0.854, 0.940 and 0.938.@*CONCLUSION@#The heterozygosities of the three loci are high and the frequencies was in accordance with Hardy-Weinberg equilibrium (P>0.05), so the three loci can be used in forensic medicine and in other genetic researches.
Subject(s)
Humans , Alleles , China/ethnology , DNA/isolation & purification , Gene Frequency , Genotype , Polymerase Chain Reaction , Polymorphism, Genetic , Tandem Repeat SequencesABSTRACT
OBJECTIVE@#To investigate the polymorphism and forensic efficiency values of STR loci D12S391/D18S865@*METHODS@#Population studies on D12S391 and D18S865 were carried out in a sample of 454 unrelated Wenzhou Han individuals using PCR followed by PAGE and silver stain detection.@*RESULTS@#11/7 alleles and 50/21 genotypes of D12S391/D18S865 were respectively observed among the 454 unrelated individuals. The genotype frequency matched the Hardy-Weinberg equilibrium, The heterozygotes (H) in D12S391/D18S865 were 0.7974/0.7379, Power of Exclusion were 0.9566/0.9077, polymorphism information content (PIC) were 0.8260/0.7279 respectively.@*CONCLUSION@#D12S391 and D18S865 were high polymorphic loci and the frequencies were in accordance with Hardy-Weinberg equilibrium (P>0.5), so the two loci can be used in forensic medicine and in other genetic research areas.