1.Relationship between the functional single nucleotide polymorphism rs1052133 in the 8-oxoguanine DNA glycosylase 1 (OGG1) gene and vitiligo in a Chinese Han population
Chao WEI ; Huini QIANG ; Tianwen GAO ; Chunying LI
Chinese Journal of Dermatology 2013;(4):227-230
Objective To estimate the relationship of the functional single nucleotide polymorphism (SNP) rs1052133 in the 8-oxoguanine DNA glycosylase 1 (OGG1) gene with vitiligo in a Chinese Han population.Methods Blood samples were collected from 800 patients with vitiligo and 800 healthy human controls,and subjected to genomic DNA extraction.PCR-restriction fragment length polymorphism (PCR-RFLP) analysis was performed to analyze the genotype of the SNP rs1052133 in the OGG1 gene.The relationship between the SNP and the risk of vitiligo was evaluated by chi-square test and unconditional logistic regression analysis.Enzyme linked immunosorbent assay (ELISA) was carried out to assess the serum level of 8-hydroxydeoxyguanosine (8-OHdG) in 83 patients with vitiligo and 83 healthy human controls,then,t test was used to compare the serum 8-OHdG level between the patients and controls.Results The frequency of CC,CG and GG genotype of the SNP rs1052133 was 16.8%,54.0% and 29.2% respectively in the patients,21.4%,52.8% and 25.8%respectively in the controls (x2 =6.26,P < 0.05).Increased frequency of G allele of the SNP rs1052133 was observed in the patients with vitiligo compared with the controls (56.2% vs.52.2%,x2 =5.16,P < 0.05).A statistically increased risk of vitiligo was associated with the CG (x2 =3.98,P < 0.05,adjusted odds ratio 1.31,95% confidence interval:1.01-1.70) and GG (x2 =6.01,P < 0.05,adjusted odds ratio 1.45,95% confidence interval:1.08-1.94) genotype of SNP rs1052133 compared with the CC genotype,which was more evident among the patients with the following characteristics:female,nonsegmental vitiligo,active vitiligo,long clinical course (> 12 months),a family history of vitiligo,and no accompanied autoimmune diseases.In addition,the patients with the CG or GG genotype of SNP rs1052133 had a higher serum 8-OHdG level than those with the CC genotype ((838.23 ± 294.11) μg/L vs.(593.84 ± 190.14) μg/L,t =3.63,P < 0.01).Conclusions The SNP rs1052133 in the OGG1 gene may be responsible for the development of vitiligo in Chinese Han populations,which is likely to be associated with defects in DNA repair.
2.Expression of a pectin lyase A gene from Aspergillus niger in Pichia pastoris GS115.
Huini QIANG ; Xinwei YANG ; Baoyu TIAN ; Chongrong KE ; Welling LIN ; Ruirui LÜ ; Wei HUANG ; Chunxiang WANG ; Jianzhong HUANG
Chinese Journal of Biotechnology 2009;25(12):1962-1968
In this study, the mature peptide sequence of a pectin lyase gene A was amplified from Aspergillus niger strain EIM-6 by using RT-PCR reverse transcription technique. The cloned gene was then inserted into a Pichia pastoris expression vector pPIC9k to produce the recombinant expression plasmid pPIC9K-pelA. By using electric shocks, we successfully transformed the recombinant pPIC9K-pelA into Pichia pastoris GS115. The activity of the engineered strain reached to 2.3 U/mL after induction with the final concentration of 1.5% methanol. SDS-PAGE analysis revealed that the pPIC9K-pelA transformant had an additional protein band of approximately 38 kD, which was not present in the control. There were no significant differences between the recombinant and native pectin lyase with regard to their hydrolysis activities.
Aspergillus niger
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enzymology
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genetics
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Electroporation
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Pichia
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genetics
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metabolism
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Polysaccharide-Lyases
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biosynthesis
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genetics
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Recombinant Proteins
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biosynthesis
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genetics
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Reverse Transcriptase Polymerase Chain Reaction