本研究旨在通过在Bac to Bac系统中表达非洲猪瘟病毒(ASFV)的P72基因,为进一步研究P72基因的结构、功能和免疫学特性奠定基础。本研究从含P72全长基因的菌株pGEX-6p-1-P72中扩增出P72基因全序列1941bp,将扩增的基因连接于pFastBac HTa...本研究旨在通过在Bac to Bac系统中表达非洲猪瘟病毒(ASFV)的P72基因,为进一步研究P72基因的结构、功能和免疫学特性奠定基础。本研究从含P72全长基因的菌株pGEX-6p-1-P72中扩增出P72基因全序列1941bp,将扩增的基因连接于pFastBac HTa杆状病毒载体中,构建重组质粒pFastBac HTa-P72,转化至感受态DH10Bac中,获得重组杆粒rBac-mid-P72,再将其转染至Sf9昆虫细胞,获得重组杆状病毒。经Western blotting和夹心ELISA分析,结果表明,P72在昆虫细胞Sf9中获得正确表达,重组P72蛋白可被特异性兔抗ASFV P72血清、P72单抗识别。结果提示,Bac to Bac系统表达的P72重组蛋白特异性强、活性稳定,具有良好的抗原性,为快速诊断ASF提供良好的候选抗原。展开更多
p53 gene plays an important role in apoptosis, which is necessary for successful invasion of trophoblast cells. The change from an arginine（Arg） to a proline（Pro） at codon 72 can influence the biological activity ...p53 gene plays an important role in apoptosis, which is necessary for successful invasion of trophoblast cells. The change from an arginine（Arg） to a proline（Pro） at codon 72 can influence the biological activity of p53, which predisposes to an increased risk of recurrent spontaneous abortion（RSA）. In order to investigate the association between p53 polymorphism at codon 72 and RSA, we conducted this meta-analysis. Pubmed, Embase and Web of science were used to identify the eligible studies. Odds ratio（OR） with 95% confidence interval（CI） was used to evaluate the strength of the association. Six studies containing 937 cases of RSA and 830 controls were included, and there was one study deviated from Hardy-Weinberg equilibrium（HWE）. There was a significant association between p53 polymorphism at codon 72 and RSA in recessive model（Pro/Pro vs. Pro/Arg＋Arg/Arg; OR=1.60, 95% CI： 1.14–2.24） and co-dominant model（Pro/Pro vs. Arg/Arg; OR=1.47, 95% CI： 1.02–2.12） whether the study that was deviated from HWE was eliminated or not. A significant association was observed in allelic model（Pro vs. Arg; OR=1.28, 95% CI： 1.04–1.57） after exclusion of the study that was deviated from HWE. No association was noted in recessive model（Pro/Pro＋Pro/Arg vs. Arg/Arg; OR=1.05, 95% CI： 0.86–1.30） and co-dominant model（Pro/Arg vs. Arg/Arg; OR=0.96, 95% CI： 0.77–1.19）. Subgroup analysis by ethnicity also indicated a significant association between p53 polymorphism at codon 72 and RSA in Caucasian group. No heterogeneity and publication bias were found. Our meta-analysis implied that p53 polymorphism at codon 72 carries high maternal risk of RSA.展开更多
文摘本研究旨在通过在Bac to Bac系统中表达非洲猪瘟病毒(ASFV)的P72基因,为进一步研究P72基因的结构、功能和免疫学特性奠定基础。本研究从含P72全长基因的菌株pGEX-6p-1-P72中扩增出P72基因全序列1941bp,将扩增的基因连接于pFastBac HTa杆状病毒载体中,构建重组质粒pFastBac HTa-P72,转化至感受态DH10Bac中,获得重组杆粒rBac-mid-P72,再将其转染至Sf9昆虫细胞,获得重组杆状病毒。经Western blotting和夹心ELISA分析,结果表明,P72在昆虫细胞Sf9中获得正确表达,重组P72蛋白可被特异性兔抗ASFV P72血清、P72单抗识别。结果提示,Bac to Bac系统表达的P72重组蛋白特异性强、活性稳定,具有良好的抗原性,为快速诊断ASF提供良好的候选抗原。
基金supported by The National Science and Technology Pillar of China program during the Twelfth Five-Year Plan Period(No.2014BAI 05B05)
文摘p53 gene plays an important role in apoptosis, which is necessary for successful invasion of trophoblast cells. The change from an arginine（Arg） to a proline（Pro） at codon 72 can influence the biological activity of p53, which predisposes to an increased risk of recurrent spontaneous abortion（RSA）. In order to investigate the association between p53 polymorphism at codon 72 and RSA, we conducted this meta-analysis. Pubmed, Embase and Web of science were used to identify the eligible studies. Odds ratio（OR） with 95% confidence interval（CI） was used to evaluate the strength of the association. Six studies containing 937 cases of RSA and 830 controls were included, and there was one study deviated from Hardy-Weinberg equilibrium（HWE）. There was a significant association between p53 polymorphism at codon 72 and RSA in recessive model（Pro/Pro vs. Pro/Arg＋Arg/Arg; OR=1.60, 95% CI： 1.14–2.24） and co-dominant model（Pro/Pro vs. Arg/Arg; OR=1.47, 95% CI： 1.02–2.12） whether the study that was deviated from HWE was eliminated or not. A significant association was observed in allelic model（Pro vs. Arg; OR=1.28, 95% CI： 1.04–1.57） after exclusion of the study that was deviated from HWE. No association was noted in recessive model（Pro/Pro＋Pro/Arg vs. Arg/Arg; OR=1.05, 95% CI： 0.86–1.30） and co-dominant model（Pro/Arg vs. Arg/Arg; OR=0.96, 95% CI： 0.77–1.19）. Subgroup analysis by ethnicity also indicated a significant association between p53 polymorphism at codon 72 and RSA in Caucasian group. No heterogeneity and publication bias were found. Our meta-analysis implied that p53 polymorphism at codon 72 carries high maternal risk of RSA.
