试验旨在研究不同品系大白猪骨形态发生蛋白受体ⅠB(bone morphogenetic protein receptor-ⅠB,BMPR-ⅠB)基因多态性及其与繁殖性状的相关性。采用Hi-SNP中高通量基因分型技术对BMPR-ⅠB基因多态位点进行分型,利用SPSS统计软件,采用最...试验旨在研究不同品系大白猪骨形态发生蛋白受体ⅠB(bone morphogenetic protein receptor-ⅠB,BMPR-ⅠB)基因多态性及其与繁殖性状的相关性。采用Hi-SNP中高通量基因分型技术对BMPR-ⅠB基因多态位点进行分型,利用SPSS统计软件,采用最小二乘法将各位点不同基因型与总产仔数、产活仔数、死胎数、畸形头数进行相关性分析。结果显示,不同品系大白猪在BMPR-ⅠB基因746A>G和852G>A位点均未检测到多态性,804G>C位点上均表现为GG基因型为优势基因型,G为优势等位基因;960C>T位点在美系大白猪中以CC基因型频率较高,而在英系大白猪中以TT基因型频率较高,在加系大白猪中CT基因型频率较高,3个不同品系大白猪群体在804G>C和960C>T位点均处于Hardy-Weinberg平衡状态(P>0.05)。804G>C位点在3个不同品系大白猪中对总产仔数、产活仔数、死胎数、畸形数等繁殖性状均无显著影响(P>0.05)。960C>T位点在美系大白猪群体中TT基因型死胎数极显著低于CC基因型1.027头(P<0.01);英系大白猪群体中TT基因型总产仔数显著高于CC基因型1.976头(P<0.05),极显著高于CT基因型2.118头(P<0.01);加系大白猪中TT基因型产活仔数显著高于CC基因型0.886头(P<0.05),死胎数、畸形头数显著低于CC基因型0.707和0.337头(P<0.05)。960C>T位点中TT基因型可作为3个不同品系大白猪繁殖性状的标记基因型。展开更多
Thermostable enterotoxinⅠ(ST1) mutant genes and thermolabile enterotoxin B subunit (LTB)genes were amplified by PCR from plasmids of Eschenichia coli C83902. The recombinantexpression plasmid pZST3LTB containing ST1-...Thermostable enterotoxinⅠ(ST1) mutant genes and thermolabile enterotoxin B subunit (LTB)genes were amplified by PCR from plasmids of Eschenichia coli C83902. The recombinantexpression plasmid pZST3LTB containing ST1-LTB fusion gene was constructed by recombinantDNA technique and then transformed into Escherichia coli BL21(DE3). The ST1-LTB fusionprotein was highly expressed in recombinant strain BL21(DE3)(pZST3LTB) and the fusionprotein was about 38.53% of total cellular protein by SDS-PAGE and thin-layer gelscanning analysis. More important, mice immunized with crude preparation containing thefusion protein inclusion bodies or inactivated recombinant strain produced antibodiesthat were able to recognize ST1 in vitro. These sera antibodies were able to neutralizethe biological activity of native ST1 in the suckling mouse assay. Hence the ST1-LTBfusion protein was nontoxic and immunogenic, the constructed recombinant strain BL21(DE3)(pZST3LTB) could be used as a candidate of vaccine strain.展开更多
为探究藏羊(Ovis aries)BMPR-ⅠB基因的序列特征、表达特征及其多态性对产羔性状的影响,本研究利用RT-qPCR、克隆测序、生物信息学方法和iMLDR技术等检测藏羊BMPR-ⅠB基因的组织表达特性,克隆了藏羊BMPR-ⅠB基因的编码区,分析了BMPR-Ⅰ...为探究藏羊(Ovis aries)BMPR-ⅠB基因的序列特征、表达特征及其多态性对产羔性状的影响,本研究利用RT-qPCR、克隆测序、生物信息学方法和iMLDR技术等检测藏羊BMPR-ⅠB基因的组织表达特性,克隆了藏羊BMPR-ⅠB基因的编码区,分析了BMPR-ⅠB理化性质和蛋白质结构,并检测了其在臧羊群体中的多态性。RT-qPCR结果表明,BMPR-ⅠB基因在下丘脑、垂体、心脏、肝脏、脾脏、肺脏、肾脏、卵巢、输卵管、子宫、瘤胃、十二指肠、臂三头肌、半腱肌、股四头肌和背最长肌等16个组织中均有表达,其中在卵巢、输卵管和子宫中的表达量高于其他组织的,在肌肉组织中的表达量最低。生物信息学分析结果表明,藏羊BMPR-ⅠB基因包含1个长为1509 bp的开放阅读框,编码502个氨基酸,其编码区氨基酸序列与绵羊的关系最近;BMPR-ⅠB蛋白的分子式为C_(2511)H_(3986)N_(694)O_(747)S_(33),分子量为56907.45 Da,总原子数为7971,理论等电点为7.78,T_(1/2)为30 h,不稳定指数为54.07,亲水性均值(grand average of hydropa-thicity,GRAVY)为-0.357;BMPR-ⅠB蛋白含信号肽,1个跨膜区,属于跨膜蛋白;存在87个潜在的磷酸化位点、2个N-糖基化修饰位点;α-螺旋、延伸链、β-转角和无规卷曲所占的比例分别为34.86%、13.15%、3.39%和48.61%,属于混合性蛋白。基因分型结果表明,藏羊BMPR-ⅠB基因编码区FecB位点存在A>G突变,且存在3种基因型,分别为野生++型、杂合突变B+型和纯合突变BB型,多态信息含量为0.07,属于低度多态,且该位点处于Hardy-Weinberg平衡状态。藏羊平均产羔数在野生++型和杂合突变+型之间差异显著(P<0.05)。本研究结果为进一步研究BMPR-ⅠB基因对藏羊繁殖性能的调控作用提供参考数据。展开更多
Objective： To comparatively study the expressive conditions of platelet activation related factors （GPⅠb, GPⅡb-Ⅲa and GMP-140） in healthy subjects and patients with coronary heart disease （CHD） of blood-stasis...Objective： To comparatively study the expressive conditions of platelet activation related factors （GPⅠb, GPⅡb-Ⅲa and GMP-140） in healthy subjects and patients with coronary heart disease （CHD） of blood-stasis （BS） or non-blood-stasis （non-BS） syndrome, and to analyze the relationship between the activities of various glycoproteins and the polymorphism of genes. Methods： With case control design adopted, patients with the CHD （40 of BS, 37 of non-BS） and 39 healthy subjects for control, all fitting to the inclusion criteria, were selected in this study. The number of affected coronary branches was recorded by the contrast examination. The mean fluorescence intensity （MFI） of GPⅠb, GPⅡb-Ⅲa, and GMP-140 （CD42b, CD61, CD62p） in patients and healthy persons was measured with flow cytometry, the polymorphism of HPA-3 gene was detected by Taqman probe technique and that of HPA-2 gene was determined by gene sequencing. Results： MFI of CD61 and CD62p was higher in the CHD patients than in the healthy control, which was also higher in patients of BS syndrome than in patients of non-BS syndrome （P〈0.05）; MFI of CD42b was lower in the CHD patients than in the healthy control （P〈0.05）, but showing insignificant difference between BS and non-BS syndrome （P〉0.05）; at the same time, no significant difference of all the above-mentioned three MFI could be found in patients with various numbers of affected coronary branches, neither in patients with different genotypes at GPⅡb HPA-3 and GPⅠb HPA-2 polymorphism loci （P〉0.05）. Conclusion： （1） The activities of GP Ⅱ b-Ⅲa and GMP-140 were obviously increased in the genesis and developing process of CHD and CHD of BS syndrome, and so they could be taken as one of the objective indexes for microscopic diagnosis of BS syndrome. （2） The level of GPⅠb was lower in CHD patients than in healthy persons, but it was not a sensitive indicator for BS syndrome of CHD. （3） Levels of GP Ⅱb-Ⅲa, GPⅠb and GMP-140 were not related with the number of affected coronary branches in CHD patients. （4） The changes in amino-acids expression induced by the two loci brought no significant influence on GPⅠb and GP Ⅱb-Ⅲa activities.展开更多
文摘试验旨在研究不同品系大白猪骨形态发生蛋白受体ⅠB(bone morphogenetic protein receptor-ⅠB,BMPR-ⅠB)基因多态性及其与繁殖性状的相关性。采用Hi-SNP中高通量基因分型技术对BMPR-ⅠB基因多态位点进行分型,利用SPSS统计软件,采用最小二乘法将各位点不同基因型与总产仔数、产活仔数、死胎数、畸形头数进行相关性分析。结果显示,不同品系大白猪在BMPR-ⅠB基因746A>G和852G>A位点均未检测到多态性,804G>C位点上均表现为GG基因型为优势基因型,G为优势等位基因;960C>T位点在美系大白猪中以CC基因型频率较高,而在英系大白猪中以TT基因型频率较高,在加系大白猪中CT基因型频率较高,3个不同品系大白猪群体在804G>C和960C>T位点均处于Hardy-Weinberg平衡状态(P>0.05)。804G>C位点在3个不同品系大白猪中对总产仔数、产活仔数、死胎数、畸形数等繁殖性状均无显著影响(P>0.05)。960C>T位点在美系大白猪群体中TT基因型死胎数极显著低于CC基因型1.027头(P<0.01);英系大白猪群体中TT基因型总产仔数显著高于CC基因型1.976头(P<0.05),极显著高于CT基因型2.118头(P<0.01);加系大白猪中TT基因型产活仔数显著高于CC基因型0.886头(P<0.05),死胎数、畸形头数显著低于CC基因型0.707和0.337头(P<0.05)。960C>T位点中TT基因型可作为3个不同品系大白猪繁殖性状的标记基因型。
文摘Thermostable enterotoxinⅠ(ST1) mutant genes and thermolabile enterotoxin B subunit (LTB)genes were amplified by PCR from plasmids of Eschenichia coli C83902. The recombinantexpression plasmid pZST3LTB containing ST1-LTB fusion gene was constructed by recombinantDNA technique and then transformed into Escherichia coli BL21(DE3). The ST1-LTB fusionprotein was highly expressed in recombinant strain BL21(DE3)(pZST3LTB) and the fusionprotein was about 38.53% of total cellular protein by SDS-PAGE and thin-layer gelscanning analysis. More important, mice immunized with crude preparation containing thefusion protein inclusion bodies or inactivated recombinant strain produced antibodiesthat were able to recognize ST1 in vitro. These sera antibodies were able to neutralizethe biological activity of native ST1 in the suckling mouse assay. Hence the ST1-LTBfusion protein was nontoxic and immunogenic, the constructed recombinant strain BL21(DE3)(pZST3LTB) could be used as a candidate of vaccine strain.
文摘为探究藏羊(Ovis aries)BMPR-ⅠB基因的序列特征、表达特征及其多态性对产羔性状的影响,本研究利用RT-qPCR、克隆测序、生物信息学方法和iMLDR技术等检测藏羊BMPR-ⅠB基因的组织表达特性,克隆了藏羊BMPR-ⅠB基因的编码区,分析了BMPR-ⅠB理化性质和蛋白质结构,并检测了其在臧羊群体中的多态性。RT-qPCR结果表明,BMPR-ⅠB基因在下丘脑、垂体、心脏、肝脏、脾脏、肺脏、肾脏、卵巢、输卵管、子宫、瘤胃、十二指肠、臂三头肌、半腱肌、股四头肌和背最长肌等16个组织中均有表达,其中在卵巢、输卵管和子宫中的表达量高于其他组织的,在肌肉组织中的表达量最低。生物信息学分析结果表明,藏羊BMPR-ⅠB基因包含1个长为1509 bp的开放阅读框,编码502个氨基酸,其编码区氨基酸序列与绵羊的关系最近;BMPR-ⅠB蛋白的分子式为C_(2511)H_(3986)N_(694)O_(747)S_(33),分子量为56907.45 Da,总原子数为7971,理论等电点为7.78,T_(1/2)为30 h,不稳定指数为54.07,亲水性均值(grand average of hydropa-thicity,GRAVY)为-0.357;BMPR-ⅠB蛋白含信号肽,1个跨膜区,属于跨膜蛋白;存在87个潜在的磷酸化位点、2个N-糖基化修饰位点;α-螺旋、延伸链、β-转角和无规卷曲所占的比例分别为34.86%、13.15%、3.39%和48.61%,属于混合性蛋白。基因分型结果表明,藏羊BMPR-ⅠB基因编码区FecB位点存在A>G突变,且存在3种基因型,分别为野生++型、杂合突变B+型和纯合突变BB型,多态信息含量为0.07,属于低度多态,且该位点处于Hardy-Weinberg平衡状态。藏羊平均产羔数在野生++型和杂合突变+型之间差异显著(P<0.05)。本研究结果为进一步研究BMPR-ⅠB基因对藏羊繁殖性能的调控作用提供参考数据。
基金Supported by the Major Program Project of National Natural Science Foundation of China(No.90409021)
文摘Objective： To comparatively study the expressive conditions of platelet activation related factors （GPⅠb, GPⅡb-Ⅲa and GMP-140） in healthy subjects and patients with coronary heart disease （CHD） of blood-stasis （BS） or non-blood-stasis （non-BS） syndrome, and to analyze the relationship between the activities of various glycoproteins and the polymorphism of genes. Methods： With case control design adopted, patients with the CHD （40 of BS, 37 of non-BS） and 39 healthy subjects for control, all fitting to the inclusion criteria, were selected in this study. The number of affected coronary branches was recorded by the contrast examination. The mean fluorescence intensity （MFI） of GPⅠb, GPⅡb-Ⅲa, and GMP-140 （CD42b, CD61, CD62p） in patients and healthy persons was measured with flow cytometry, the polymorphism of HPA-3 gene was detected by Taqman probe technique and that of HPA-2 gene was determined by gene sequencing. Results： MFI of CD61 and CD62p was higher in the CHD patients than in the healthy control, which was also higher in patients of BS syndrome than in patients of non-BS syndrome （P〈0.05）; MFI of CD42b was lower in the CHD patients than in the healthy control （P〈0.05）, but showing insignificant difference between BS and non-BS syndrome （P〉0.05）; at the same time, no significant difference of all the above-mentioned three MFI could be found in patients with various numbers of affected coronary branches, neither in patients with different genotypes at GPⅡb HPA-3 and GPⅠb HPA-2 polymorphism loci （P〉0.05）. Conclusion： （1） The activities of GP Ⅱ b-Ⅲa and GMP-140 were obviously increased in the genesis and developing process of CHD and CHD of BS syndrome, and so they could be taken as one of the objective indexes for microscopic diagnosis of BS syndrome. （2） The level of GPⅠb was lower in CHD patients than in healthy persons, but it was not a sensitive indicator for BS syndrome of CHD. （3） Levels of GP Ⅱb-Ⅲa, GPⅠb and GMP-140 were not related with the number of affected coronary branches in CHD patients. （4） The changes in amino-acids expression induced by the two loci brought no significant influence on GPⅠb and GP Ⅱb-Ⅲa activities.
