BACKGROUND: Budd-Chiari syndrome (BCS) is a type of disease characterized by portal hypertension and/or hy- pertension of the inferior vena cava (IVC) due to the ob- struction of the hepatic veins (HV) and/or intrahep...BACKGROUND: Budd-Chiari syndrome (BCS) is a type of disease characterized by portal hypertension and/or hy- pertension of the inferior vena cava (IVC) due to the ob- struction of the hepatic veins (HV) and/or intrahepatic IVC outlet. Being etiologically complicated and obscure, BCS can be acquired or idiopathic and several gene muta- tions may be contributable. This study was to explore whether prothrombin gene mutation (F G20210A) takes part in the pathogenesis of BCS and to investigate their cor- relativity. METHODS: In 38 proven BCS patients and 70 controls, polymerase chain reaction-restriction fragment length poly- morphism (PCR-RFLP) was used to find F G20210A mutation. To detect whether there are any mutations, four steps were taken: purification of genome DNA from whole blood, amplification of special fragment by polymerase chain reaction, digestion of the fragment via restriction en- donuclease, and analysis of results by polyacrylamide gel electrophoresis. RESULTS: F G20210A mutation was not detected in all patients and controls. CONCLUSIONS: No F G20210A mutation exists in Chi- nese patients with BCS, nor correlativity between the oc- currence of BCS and F G20210A mutation. The etiology of BCS in the Chinese needs further investigation.展开更多
Stripe rust, caused by Puccinia striiformis f. sp. tritici （Pst）, is one of the most widespread and destructive wheat diseases in many wheat-growing regions of the world. The winter wheat translocation line H9014-14...Stripe rust, caused by Puccinia striiformis f. sp. tritici （Pst）, is one of the most widespread and destructive wheat diseases in many wheat-growing regions of the world. The winter wheat translocation line H9014-14-4-6-1 has all stage resistance. To identify stripe rust resistance genes, the segregating populations were developed from the cross between H9014-14-4-6-1 and Mingxian 169 （a wheat cultivar susceptible to all Pst races identified in China）. The seedlings of the parents and F1 plants, Fz, F3 and BC1 generations were tested with Pst races under controlled greenhouse conditions. Two genes for resistance to stripe rust were identified, one dominant gene conferred resistance to SUN11-4, temporarily designated YrH9014 and the other recessive gene conferred resistance to CYR33. The bulked segregant analysis and simple sequence repeat （SSR） markers were used to identify polymorphic markers associated with YrH9014. Seven polymorphic SSR markers were used to genotype the F2 population inoculated with SUN11-4. A linkage map was constructed according to the genotypes of seven SSR markers and resistance gene. The molecular map spanned 24.3 cM, and the genetic distance of the two closest markers Xbarc13 and Xbarc55 to gene locus was 1.4 and 3.6 cM, respectively. Based on the position of SSR marker, the resistance gene YrH9014 was located on chromosome arm 2BS. Amplification of a set of nulli-tetrasomic Chinese Spring lines with SSR marker Xbarc13 indicated that YrH9014 was located on chromosome 2B. Based on chromosomal location, the reaction patterns and pedigree analysis, YrH9014 should be a novel resistance gene to stripe rust. This new gene and flanking markers got from this study should be useful for marker-assisted selection （MAS） in breeding programs for stripe rust.展开更多
扩增禽4型副黏病毒(Avian paramyxovirus type 4,APMV-4)新疆野鸭源分离株APMV-4/Pintail/CH(XJ)/01/2016的F基因,并构建原核表达载体。参考GenBank上公布的APMV-4序列,通过DNAStar设计F基因特异性引物,以APMV-4/Pintail/CH(XJ)/01/201...扩增禽4型副黏病毒(Avian paramyxovirus type 4,APMV-4)新疆野鸭源分离株APMV-4/Pintail/CH(XJ)/01/2016的F基因,并构建原核表达载体。参考GenBank上公布的APMV-4序列,通过DNAStar设计F基因特异性引物,以APMV-4/Pintail/CH(XJ)/01/2016基因组RNA为模板,采用RT-PCR方法扩增F基因编码区,连接于pMD19-T载体,用BamHⅠ和HindⅢ酶切原核表达载体pET-30a重组质粒,将酶切产物克隆至pET-30a多克隆位点上,转化至大肠杆菌DH5α感受态细胞中,构建F基因原核表达载体,并对阳性重组质粒进行PCR和测序鉴定。结果显示,扩增出的片段大小为1701 bp,经测序鉴定该扩增片段为F基因全长,与GenBank中收录的APMV-4序列核苷酸相似性为99%;连接表达载体结果表明,原核表达载体pET-30a-F成功构建,为APMV-4诊断方法的建立提供了良好的技术基础。展开更多
Stripe rust is one of the most important diseases of wheat worldwide. Inheritance of stripe rust resistance and mapping of resistance gene with simple sequence repeat （SSR） markers are studied to formulate efficient...Stripe rust is one of the most important diseases of wheat worldwide. Inheritance of stripe rust resistance and mapping of resistance gene with simple sequence repeat （SSR） markers are studied to formulate efficient strategies for breeding cultivars resistant to stripe rust. Zhongliang 88375, a common wheat line, is highly resistant to all three rusts of wheat in China. The gene conferring rust disease was deduced originating from Elytrigia intermedium. Genetic analysis of Zhongliang 88375 indicated that the resistance to PST race CYR31 was controlled by a single dominant gene, temporarily designated as Yr88375. To molecular map Yr88375, a F2 segregating population consisting of 163 individuals was constructed on the basis of the hybridization between Zhongliang 88375 and a susceptible wheat line Mingxian 169; 320 SSR primer pairs were used for analyzing the genetic linkage relation. Six SSR markers, Xgwm335, Xwmc289, Xwmc810, Xgdmll6, Xbarc59, and Xwmc783, are linked to Yr88375 as they were all located on chromosome 5BL Yr88375 was also located on that chromosome arm, closely linked to Xgdmll6 and Xwmc810 with genetic distances of 3.1 and 3.9 cM, respectively. The furthest marker Xwmc783 was 13.5 cM to Yr88375. Hence, pedigree analysis of Zhongliang 88375 combined with SSR markers supports the conclusion that the highly resistance gene Yr88375 derived from Elytrigia intermedium is a novel gene for resistance to stripe rust in wheat. It could play an important role in wheat breeding programs for stripe rust resistance.展开更多
[Objective] This study aimed to construct the recombinant expression plasmids containing H and F protein genes of Canine distemper virus isolated from a mink and to express these two genes in prekaryotic cells as well...[Objective] This study aimed to construct the recombinant expression plasmids containing H and F protein genes of Canine distemper virus isolated from a mink and to express these two genes in prekaryotic cells as well as to study the reactogenieity of the expressed products. [ Method ] RT-PCR amplification was used to obtain H and F protein genes; TA cloning and subclonlng techniques were used to construct the cloning plasmids（pMD-18T-H and pMD-18T-F） and recombinant expression plasmids（pET28a-H and pET28a-F） ; SDS-PAGE and Western-blotting were adopted to verify whether the target proteins were successfully expressed. [ Result] The recombinant expression plasmids pET28a-H and pET28a-F containing H and F protein genes of Canine distemper virus isolated from a mink were successfully constructed, and both the expressed H and F proteins with respectively relative molecular mass of 31 400 and 38 200 produced positive reac- tion with the CDV standard positive serum. [ Conclusion] The H and F proteins expressed in prokaryotic cells were the same with the natural ones in terms of reac- togenicity, which can be utilized for diagnosis of a CDV＇s infection or for an epidemiological investigation. Meanwhile, they also provide a basis for developing ge- netically engineered subunit vaccines.展开更多
supported by the Earmarked Fund for the Modern Agroindustry Technology Research System, China （CARS-41）;the National High Technology Research and Development Program of China （2011AA100301）
Stripe rust （yellow rust）, caused by Puccinia striiformis f. sp. tritici （Pst）, is one of the most devastating diseases of wheat throughout the world. H9020-1-6-8-3 is a translocation line originally developed fro...Stripe rust （yellow rust）, caused by Puccinia striiformis f. sp. tritici （Pst）, is one of the most devastating diseases of wheat throughout the world. H9020-1-6-8-3 is a translocation line originally developed from interspeciifc hybridization between wheat line 7182 and Psathyrostachys huashanica Keng and is resistant to most Pst races in China. To identify the resistance gene（s） in the translocation line, H9020-1-6-8-3 was crossed with susceptible cultivar Mingxian 169, and seedlings of the parents, F1, F2, F3, and BC1 generations were tested with prevalent Chinese Pst race CYR32 under controlled greenhouse conditions. The results indicated that there is a single dominant gene, temporarily designated as YrH9020a, conferring resistance to CYR32. The resistance gene was mapped by the F2 population from Mingxian 169/H9020-1-6-8-3. It was linked to six microsatellite markers, including Xbarc196, Xbarc202, Xbarc96, Xgpw4372, Xbarc21, and Xgdm141, lfanked by Xbarc96 and Xbarc202 with at 4.5 and 8.3 cM, respectively. Based on the chromosomal locations of these markers and the test of Chinese Spring （CS） nullitetrasomic and ditelosomic lines, the gene was assigned to chromosome 6D. According to the origin and the chromosomal location, YrH9020a might be a new resistance gene to stripe rust. The lfanking markers linked to YrH9020a could be useful for marker-assisted selection in breeding programs.展开更多
基金This study was partly supported by a grant from the Science and TechnologyKey Project of Henan Province, China ( No. 0224630176 ).
文摘BACKGROUND: Budd-Chiari syndrome (BCS) is a type of disease characterized by portal hypertension and/or hy- pertension of the inferior vena cava (IVC) due to the ob- struction of the hepatic veins (HV) and/or intrahepatic IVC outlet. Being etiologically complicated and obscure, BCS can be acquired or idiopathic and several gene muta- tions may be contributable. This study was to explore whether prothrombin gene mutation (F G20210A) takes part in the pathogenesis of BCS and to investigate their cor- relativity. METHODS: In 38 proven BCS patients and 70 controls, polymerase chain reaction-restriction fragment length poly- morphism (PCR-RFLP) was used to find F G20210A mutation. To detect whether there are any mutations, four steps were taken: purification of genome DNA from whole blood, amplification of special fragment by polymerase chain reaction, digestion of the fragment via restriction en- donuclease, and analysis of results by polyacrylamide gel electrophoresis. RESULTS: F G20210A mutation was not detected in all patients and controls. CONCLUSIONS: No F G20210A mutation exists in Chi- nese patients with BCS, nor correlativity between the oc- currence of BCS and F G20210A mutation. The etiology of BCS in the Chinese needs further investigation.
基金supported by the 111 Project from the Education Ministry of China(B07049)the National 11th Five-Year Plan Key Project(2006BAD08A05)Toxicity Variation of Wheat Stripe Rust Pathogen and Demonstration of Integrated Management of Stripe Rust, China (200903035-02)
文摘Stripe rust, caused by Puccinia striiformis f. sp. tritici （Pst）, is one of the most widespread and destructive wheat diseases in many wheat-growing regions of the world. The winter wheat translocation line H9014-14-4-6-1 has all stage resistance. To identify stripe rust resistance genes, the segregating populations were developed from the cross between H9014-14-4-6-1 and Mingxian 169 （a wheat cultivar susceptible to all Pst races identified in China）. The seedlings of the parents and F1 plants, Fz, F3 and BC1 generations were tested with Pst races under controlled greenhouse conditions. Two genes for resistance to stripe rust were identified, one dominant gene conferred resistance to SUN11-4, temporarily designated YrH9014 and the other recessive gene conferred resistance to CYR33. The bulked segregant analysis and simple sequence repeat （SSR） markers were used to identify polymorphic markers associated with YrH9014. Seven polymorphic SSR markers were used to genotype the F2 population inoculated with SUN11-4. A linkage map was constructed according to the genotypes of seven SSR markers and resistance gene. The molecular map spanned 24.3 cM, and the genetic distance of the two closest markers Xbarc13 and Xbarc55 to gene locus was 1.4 and 3.6 cM, respectively. Based on the position of SSR marker, the resistance gene YrH9014 was located on chromosome arm 2BS. Amplification of a set of nulli-tetrasomic Chinese Spring lines with SSR marker Xbarc13 indicated that YrH9014 was located on chromosome 2B. Based on chromosomal location, the reaction patterns and pedigree analysis, YrH9014 should be a novel resistance gene to stripe rust. This new gene and flanking markers got from this study should be useful for marker-assisted selection （MAS） in breeding programs for stripe rust.
文摘扩增禽4型副黏病毒(Avian paramyxovirus type 4,APMV-4)新疆野鸭源分离株APMV-4/Pintail/CH(XJ)/01/2016的F基因,并构建原核表达载体。参考GenBank上公布的APMV-4序列,通过DNAStar设计F基因特异性引物,以APMV-4/Pintail/CH(XJ)/01/2016基因组RNA为模板,采用RT-PCR方法扩增F基因编码区,连接于pMD19-T载体,用BamHⅠ和HindⅢ酶切原核表达载体pET-30a重组质粒,将酶切产物克隆至pET-30a多克隆位点上,转化至大肠杆菌DH5α感受态细胞中,构建F基因原核表达载体,并对阳性重组质粒进行PCR和测序鉴定。结果显示,扩增出的片段大小为1701 bp,经测序鉴定该扩增片段为F基因全长,与GenBank中收录的APMV-4序列核苷酸相似性为99%;连接表达载体结果表明,原核表达载体pET-30a-F成功构建,为APMV-4诊断方法的建立提供了良好的技术基础。
基金the National 973 Programof China(G2000016200)Program for Changjiang Scholars and Innovative Research Teamin University from Ministry of Education of China(200558)
文摘Stripe rust is one of the most important diseases of wheat worldwide. Inheritance of stripe rust resistance and mapping of resistance gene with simple sequence repeat （SSR） markers are studied to formulate efficient strategies for breeding cultivars resistant to stripe rust. Zhongliang 88375, a common wheat line, is highly resistant to all three rusts of wheat in China. The gene conferring rust disease was deduced originating from Elytrigia intermedium. Genetic analysis of Zhongliang 88375 indicated that the resistance to PST race CYR31 was controlled by a single dominant gene, temporarily designated as Yr88375. To molecular map Yr88375, a F2 segregating population consisting of 163 individuals was constructed on the basis of the hybridization between Zhongliang 88375 and a susceptible wheat line Mingxian 169; 320 SSR primer pairs were used for analyzing the genetic linkage relation. Six SSR markers, Xgwm335, Xwmc289, Xwmc810, Xgdmll6, Xbarc59, and Xwmc783, are linked to Yr88375 as they were all located on chromosome 5BL Yr88375 was also located on that chromosome arm, closely linked to Xgdmll6 and Xwmc810 with genetic distances of 3.1 and 3.9 cM, respectively. The furthest marker Xwmc783 was 13.5 cM to Yr88375. Hence, pedigree analysis of Zhongliang 88375 combined with SSR markers supports the conclusion that the highly resistance gene Yr88375 derived from Elytrigia intermedium is a novel gene for resistance to stripe rust in wheat. It could play an important role in wheat breeding programs for stripe rust resistance.
基金Supported by the Natural Science Foundation of Jilin Province(201115194)Education Department of Jilin Province(2009.No.66)
文摘[Objective] This study aimed to construct the recombinant expression plasmids containing H and F protein genes of Canine distemper virus isolated from a mink and to express these two genes in prekaryotic cells as well as to study the reactogenieity of the expressed products. [ Method ] RT-PCR amplification was used to obtain H and F protein genes; TA cloning and subclonlng techniques were used to construct the cloning plasmids（pMD-18T-H and pMD-18T-F） and recombinant expression plasmids（pET28a-H and pET28a-F） ; SDS-PAGE and Western-blotting were adopted to verify whether the target proteins were successfully expressed. [ Result] The recombinant expression plasmids pET28a-H and pET28a-F containing H and F protein genes of Canine distemper virus isolated from a mink were successfully constructed, and both the expressed H and F proteins with respectively relative molecular mass of 31 400 and 38 200 produced positive reac- tion with the CDV standard positive serum. [ Conclusion] The H and F proteins expressed in prokaryotic cells were the same with the natural ones in terms of reac- togenicity, which can be utilized for diagnosis of a CDV＇s infection or for an epidemiological investigation. Meanwhile, they also provide a basis for developing ge- netically engineered subunit vaccines.
基金supported by the Earmarked Fund for the Modern Agroindustry Technology Research System, China (CARS-41)the National High Technology Research and Development Program of China (2011AA100301)
文摘supported by the Earmarked Fund for the Modern Agroindustry Technology Research System, China （CARS-41）;the National High Technology Research and Development Program of China （2011AA100301）
基金supported by the Programme of Introducing Talents of Discipline to Universities, Ministry of Education, China (111 Project, B07049)the National Basic Research Program of China (973 Program, 2013CB127700)+2 种基金the Science and Technology Co-ordinating Innovative Engineering Project of Shaanxi Province, China (2012KTCL02-10)the National Natural Science Foundation of China (30771397)the China Postdoctoral Science Foundation (2012M512034)
文摘Stripe rust （yellow rust）, caused by Puccinia striiformis f. sp. tritici （Pst）, is one of the most devastating diseases of wheat throughout the world. H9020-1-6-8-3 is a translocation line originally developed from interspeciifc hybridization between wheat line 7182 and Psathyrostachys huashanica Keng and is resistant to most Pst races in China. To identify the resistance gene（s） in the translocation line, H9020-1-6-8-3 was crossed with susceptible cultivar Mingxian 169, and seedlings of the parents, F1, F2, F3, and BC1 generations were tested with prevalent Chinese Pst race CYR32 under controlled greenhouse conditions. The results indicated that there is a single dominant gene, temporarily designated as YrH9020a, conferring resistance to CYR32. The resistance gene was mapped by the F2 population from Mingxian 169/H9020-1-6-8-3. It was linked to six microsatellite markers, including Xbarc196, Xbarc202, Xbarc96, Xgpw4372, Xbarc21, and Xgdm141, lfanked by Xbarc96 and Xbarc202 with at 4.5 and 8.3 cM, respectively. Based on the chromosomal locations of these markers and the test of Chinese Spring （CS） nullitetrasomic and ditelosomic lines, the gene was assigned to chromosome 6D. According to the origin and the chromosomal location, YrH9020a might be a new resistance gene to stripe rust. The lfanking markers linked to YrH9020a could be useful for marker-assisted selection in breeding programs.
