根据已发表的牛流产型布鲁氏菌(B.abrotus)HtrA(High temperature requirment A)基因的核酸序列设计引物,从新疆绵羊种布鲁氏菌(B.ovis)基因组中扩增到了大约1600bp的片段。将该片段纯化后克隆到PBS-T载体上,对所得到的重组质粒进行PCR...根据已发表的牛流产型布鲁氏菌(B.abrotus)HtrA(High temperature requirment A)基因的核酸序列设计引物,从新疆绵羊种布鲁氏菌(B.ovis)基因组中扩增到了大约1600bp的片段。将该片段纯化后克隆到PBS-T载体上,对所得到的重组质粒进行PCR鉴定、酶切分析后,对克隆的片段进行测序表明,新疆绵羊种布鲁氏菌HtrA基因与发表的牛流产型布鲁氏菌、马耳他布鲁氏菌(B.melitensis)、猪种布鲁氏菌(B.suis)的HtrA基因核苷酸序列的同源性分别为99.68%、99.81%、99.55%,推导的氨基酸序列也存在很高的同源性。展开更多
目的探讨高温需要蛋白A(htrA)和酪蛋白裂解酶P(clpP)单基因缺失对变异链球菌铁代谢的影响。方法采用浊度法测定变异链球菌标准株、htrA缺陷株和clpP缺陷株在0.1、0.2及0.3mmol/L铁限制条件培养基中培养24h的吸光度(A)值,并测定加入外源...目的探讨高温需要蛋白A(htrA)和酪蛋白裂解酶P(clpP)单基因缺失对变异链球菌铁代谢的影响。方法采用浊度法测定变异链球菌标准株、htrA缺陷株和clpP缺陷株在0.1、0.2及0.3mmol/L铁限制条件培养基中培养24h的吸光度(A)值,并测定加入外源性铁或标准株分别与2种缺陷株在0.3mmol/L铁限制培养基中混合培养24h后的A值。结果与标准株相比,htrA缺陷株在浓度0.3mmol/L时A值差异有统计学意义(P<0.01);clpP缺陷株在0.2mmol/L时A值差异即有统计学意义(P<0.05),并且其值随铁螯合剂浓度的增加而减小。加入FeSO4培养24h,htrA缺陷株与标准株A值差异无统计学意义(P>0.05);clpP缺陷株的A值低于标准株(1.002±0.014 vs 1.325±0.010,P<0.05)。与标准株按1∶1比例混合培养24h,htrA缺陷株与其单独生长时的A值的差异无统计学意义(P>0.05);clpP缺陷株的A值高于其单独培养时的A值(10.878±0.005 vs 0.664±0.006),差异有统计学意义(P<0.05)。结论 htrA和clpP基因分别在一定程度上影响变异链球菌的铁代谢。展开更多
Background The present study was undertaken to replicate the associations of representative polymorphisms in three genes （complement factor H （CFH）, complement factor B （BF） and HtrA serine peptidase 1 （HTRA1）...Background The present study was undertaken to replicate the associations of representative polymorphisms in three genes （complement factor H （CFH）, complement factor B （BF） and HtrA serine peptidase 1 （HTRA1）） with exudative age-related macular degeneration （AMD） in a Han Chinese population, and to test if the modifiable environmental factors affect AMD susceptibility associated with different type of genotype in these genes. Methods An age, gender and ethnicity matched case-control study was conducted to genotype the representative single neucleotide polymorphisms （SNPs） loci including rs1061170 and rs1410996 in CFH, rs641153 and rs4151667 in BF and rs11200638 in HTRA1 gene in 144 exudative AMD patients and 126 normal controls using PCR-RFLP and direct resequencing. The demographic characteristics and behavioral risk factors were also recorded. Allelic and genotypic associations for individual SNP and joint associations with two loci were performed. The gene-gene and gene-environment interactions were analyzed using multivariate non-conditional Logistic regression analysis. Results The C risk allele frequencies for CFH Y402H （rs1061170） in cases and controls were 12.5% and 5.4% respectively, which were much lower than those in Caucasians （P 〈0.001）. Compared with TT homozygous genotype, the CT heterozygous genotype was positively associated with AMD with odds ratio （OR） of 3.23 （1.36-5.07）. However, the population attributable risk （PAR） of C allele was only 3.3% （1.4%-4.3%）. rs1410996 was also associated with AMD independent of Y402H. The ORs of exudative AMD for individuals carrying one copy risk allele and two copy risk alleles were 2.57 （1.21-5.45） and 4.76 （2.15-10.55） respectively, with correspondent PARs of 28.3% （2.0%-40.5%） and 38.2% （21.8%-45.4%）. rs11200636 in HTRA1 was another susceptible locus for AMD and the risk homozygotes were significantly susceptible for exudutive AMD （OR=3.98, 1.88-8.43） with PAR of 38.9% （24.3%-45.8%）. Education status and cigarette smoking were also related to exudative AMD. After controlling for environmental risk factors, CFH and HTRA1 SNPs were independently associated with exudative AMD, with OR of 3.50 （1.45-8.45） for CT genotype in Y402H, 3.34 （1.33-8.36） for GG genotype in rs1410996 and 3.85 （1.58-9.42） for AA genotype in rs11200638 respectively. The interaction analysis between gene and environmental factors showed that smoking synergistically increased susceptibility of AMD for heterozygotes of rs1410996, with ORinteraction of 7.33 （Pinteraction=0.029）. Conclusions In a Han Chinese population, CFH and HTRA1 polymorphisms appear to be independently and possibly additively hereditary contributors to exudative AMD. Y402H polymorphism conferred a significant but relatively lower contribution in Chinese than in Caucasians with a low frequency of risk allele. The gene-environment interaction may be a best way to encourage those with a high genetic risk to prevent AMD by avoiding modifiable factors until there is effective treatment for AMD.展开更多
文摘根据已发表的牛流产型布鲁氏菌(B.abrotus)HtrA(High temperature requirment A)基因的核酸序列设计引物,从新疆绵羊种布鲁氏菌(B.ovis)基因组中扩增到了大约1600bp的片段。将该片段纯化后克隆到PBS-T载体上,对所得到的重组质粒进行PCR鉴定、酶切分析后,对克隆的片段进行测序表明,新疆绵羊种布鲁氏菌HtrA基因与发表的牛流产型布鲁氏菌、马耳他布鲁氏菌(B.melitensis)、猪种布鲁氏菌(B.suis)的HtrA基因核苷酸序列的同源性分别为99.68%、99.81%、99.55%,推导的氨基酸序列也存在很高的同源性。
文摘目的探讨高温需要蛋白A(htrA)和酪蛋白裂解酶P(clpP)单基因缺失对变异链球菌铁代谢的影响。方法采用浊度法测定变异链球菌标准株、htrA缺陷株和clpP缺陷株在0.1、0.2及0.3mmol/L铁限制条件培养基中培养24h的吸光度(A)值,并测定加入外源性铁或标准株分别与2种缺陷株在0.3mmol/L铁限制培养基中混合培养24h后的A值。结果与标准株相比,htrA缺陷株在浓度0.3mmol/L时A值差异有统计学意义(P<0.01);clpP缺陷株在0.2mmol/L时A值差异即有统计学意义(P<0.05),并且其值随铁螯合剂浓度的增加而减小。加入FeSO4培养24h,htrA缺陷株与标准株A值差异无统计学意义(P>0.05);clpP缺陷株的A值低于标准株(1.002±0.014 vs 1.325±0.010,P<0.05)。与标准株按1∶1比例混合培养24h,htrA缺陷株与其单独生长时的A值的差异无统计学意义(P>0.05);clpP缺陷株的A值高于其单独培养时的A值(10.878±0.005 vs 0.664±0.006),差异有统计学意义(P<0.05)。结论 htrA和clpP基因分别在一定程度上影响变异链球菌的铁代谢。
文摘Background The present study was undertaken to replicate the associations of representative polymorphisms in three genes （complement factor H （CFH）, complement factor B （BF） and HtrA serine peptidase 1 （HTRA1）） with exudative age-related macular degeneration （AMD） in a Han Chinese population, and to test if the modifiable environmental factors affect AMD susceptibility associated with different type of genotype in these genes. Methods An age, gender and ethnicity matched case-control study was conducted to genotype the representative single neucleotide polymorphisms （SNPs） loci including rs1061170 and rs1410996 in CFH, rs641153 and rs4151667 in BF and rs11200638 in HTRA1 gene in 144 exudative AMD patients and 126 normal controls using PCR-RFLP and direct resequencing. The demographic characteristics and behavioral risk factors were also recorded. Allelic and genotypic associations for individual SNP and joint associations with two loci were performed. The gene-gene and gene-environment interactions were analyzed using multivariate non-conditional Logistic regression analysis. Results The C risk allele frequencies for CFH Y402H （rs1061170） in cases and controls were 12.5% and 5.4% respectively, which were much lower than those in Caucasians （P 〈0.001）. Compared with TT homozygous genotype, the CT heterozygous genotype was positively associated with AMD with odds ratio （OR） of 3.23 （1.36-5.07）. However, the population attributable risk （PAR） of C allele was only 3.3% （1.4%-4.3%）. rs1410996 was also associated with AMD independent of Y402H. The ORs of exudative AMD for individuals carrying one copy risk allele and two copy risk alleles were 2.57 （1.21-5.45） and 4.76 （2.15-10.55） respectively, with correspondent PARs of 28.3% （2.0%-40.5%） and 38.2% （21.8%-45.4%）. rs11200636 in HTRA1 was another susceptible locus for AMD and the risk homozygotes were significantly susceptible for exudutive AMD （OR=3.98, 1.88-8.43） with PAR of 38.9% （24.3%-45.8%）. Education status and cigarette smoking were also related to exudative AMD. After controlling for environmental risk factors, CFH and HTRA1 SNPs were independently associated with exudative AMD, with OR of 3.50 （1.45-8.45） for CT genotype in Y402H, 3.34 （1.33-8.36） for GG genotype in rs1410996 and 3.85 （1.58-9.42） for AA genotype in rs11200638 respectively. The interaction analysis between gene and environmental factors showed that smoking synergistically increased susceptibility of AMD for heterozygotes of rs1410996, with ORinteraction of 7.33 （Pinteraction=0.029）. Conclusions In a Han Chinese population, CFH and HTRA1 polymorphisms appear to be independently and possibly additively hereditary contributors to exudative AMD. Y402H polymorphism conferred a significant but relatively lower contribution in Chinese than in Caucasians with a low frequency of risk allele. The gene-environment interaction may be a best way to encourage those with a high genetic risk to prevent AMD by avoiding modifiable factors until there is effective treatment for AMD.
