[ Objective] The study was to clone HN and F genes from GX1 strain of goose paramyxovirus and analyze their sequences. [ Method] According to the full nucleotide sequence of GPV-SF02 strain of goose paramyxovirus, two...[ Objective] The study was to clone HN and F genes from GX1 strain of goose paramyxovirus and analyze their sequences. [ Method] According to the full nucleotide sequence of GPV-SF02 strain of goose paramyxovirus, two pairs of pdmers were designed to amplify the HN and F genes from GX1 strain of goose paramyxovirus isolated from diseased goose in Guangxi Zhuang Autonomous Region; the amplified products were ligated into pMD18-T vector and sequenced. [ Result ] HN and F genes of this strain tested were 1 716 and 1 662 bp in full nucleotide length, respectively; both showed the homologues of about 97.3% with GPV- SF02 strain, of 80.3% -97.5% with strains LaSota, F48E9 and JS, of just 84.8% with Miyadera strain. [ Conclusion] The results show that isolated strain BX1 matches to virulent APMV-1 strain, belonging to genotype Ⅶ of APMV-1 strain.展开更多
BACKGROUND Dilated cardiomyopathy(DCM)is a genetically heterogeneous cardiac disorder characterized by left ventricular dilation and contractile dysfunction.The substantial genetic heterogeneity evident in patients wi...BACKGROUND Dilated cardiomyopathy(DCM)is a genetically heterogeneous cardiac disorder characterized by left ventricular dilation and contractile dysfunction.The substantial genetic heterogeneity evident in patients with DCM contributes to variable disease severity and complicates overall prognosis,which can be very poor.AIM To identify pathogenic genes in DCM through pedigree analysis.METHODS Our research team identified a patient with DCM in the clinic.Through invest-igation,we found that the family of this patient has a typical DCM pedigree.High-throughput sequencing technology,next-generation sequencing,was used to sequence the whole exomes of seven samples in the pedigree.RESULTS A novel and potentially pathogenic gene mutation-ANK2p.F3067L-was discovered.The mutation was completely consistent with the clinical information for this DCM pedigree.Sanger sequencing was used to further verify the locus of the mutation in pedigree samples.These results were consistent with those of high-throughput sequencing.CONCLUSIONS ANK2p.F3067L is considered a novel and potentially pathogenic gene mutation in DCM.展开更多
将新城疫病毒 (NDV)长春株和四平株 HN插入 p IRES1多克隆位点 (Eco R )中 ,构建成核酸表达疫苗 p IRc HN和 p IRs HN,然后切除 p IRc HN和 p IRs HN的新霉素基因 ,将长春株和四平株 F基因分别插入其中 ,构建成 p IRc HNF和 p IRs HNF,...将新城疫病毒 (NDV)长春株和四平株 HN插入 p IRES1多克隆位点 (Eco R )中 ,构建成核酸表达疫苗 p IRc HN和 p IRs HN,然后切除 p IRc HN和 p IRs HN的新霉素基因 ,将长春株和四平株 F基因分别插入其中 ,构建成 p IRc HNF和 p IRs HNF,而后分别转染 Hela细胞。经血凝效价测定、Western blot分析 ,弱毒株构建的核酸疫苗的血凝活性比强毒株高 1个数量级 ,Hela细胞表达的 HN蛋白量以 p IRc HN最高 ,p IRs HN次之 ,p IRc HNF和 p IRs HNF较低。将重组疫苗转染 Hela细胞 ,用兔抗鸡 Ig Y进行间接免疫荧光试验 ,结果 ,在细胞膜和细胞浆中观察到了特异性的黄绿色荧光 ,证明表达产物具有特异性。展开更多
Virion RNA was abstracted from purified type I Avian paramyxovirus strain YN-PA01(isolated from parrot)and used as a template. The fragment containing the fusion gene(F) and hemagglutinin-neuraminidase gene(HN) of the...Virion RNA was abstracted from purified type I Avian paramyxovirus strain YN-PA01(isolated from parrot)and used as a template. The fragment containing the fusion gene(F) and hemagglutinin-neuraminidase gene(HN) of the isolate was amplified by RT-PCR and cloned to the pMD18-T Vector. Using primer walking method the complete sequence of F-HN genes was obtained finally.And the respective amino acid sequence was deduced. Through relative software the phylogenetic trees on F gene and HN gene were constructed between strain YN-PA01 and reference strains. The results showed that strain YN-PA01 comparing with reference strains displays 98.7%-83.2% nucleotide homology and 98.1%-86.2% amino acid homology on F gene; 97.4%-79.1% nucleotide homology and 97.2%-83.2% amino acid homology on HN gene. Additional 6 amino acids are encoded by the HN gene ORF of strain YN-PA01 comparing with national reference strains.The studied strain YN-PA01 exhibits highest identities with strain JS/5[O1/Go either analyzed on F gene or HN gene.展开更多
基金Supported by the Development Program for Guangxi Science andTechnology(0719004-3G)~~
文摘[ Objective] The study was to clone HN and F genes from GX1 strain of goose paramyxovirus and analyze their sequences. [ Method] According to the full nucleotide sequence of GPV-SF02 strain of goose paramyxovirus, two pairs of pdmers were designed to amplify the HN and F genes from GX1 strain of goose paramyxovirus isolated from diseased goose in Guangxi Zhuang Autonomous Region; the amplified products were ligated into pMD18-T vector and sequenced. [ Result ] HN and F genes of this strain tested were 1 716 and 1 662 bp in full nucleotide length, respectively; both showed the homologues of about 97.3% with GPV- SF02 strain, of 80.3% -97.5% with strains LaSota, F48E9 and JS, of just 84.8% with Miyadera strain. [ Conclusion] The results show that isolated strain BX1 matches to virulent APMV-1 strain, belonging to genotype Ⅶ of APMV-1 strain.
基金Supported by the Jilin Provincial Healthcare Talent Special Program,No.2019SCZT08.
文摘BACKGROUND Dilated cardiomyopathy(DCM)is a genetically heterogeneous cardiac disorder characterized by left ventricular dilation and contractile dysfunction.The substantial genetic heterogeneity evident in patients with DCM contributes to variable disease severity and complicates overall prognosis,which can be very poor.AIM To identify pathogenic genes in DCM through pedigree analysis.METHODS Our research team identified a patient with DCM in the clinic.Through invest-igation,we found that the family of this patient has a typical DCM pedigree.High-throughput sequencing technology,next-generation sequencing,was used to sequence the whole exomes of seven samples in the pedigree.RESULTS A novel and potentially pathogenic gene mutation-ANK2p.F3067L-was discovered.The mutation was completely consistent with the clinical information for this DCM pedigree.Sanger sequencing was used to further verify the locus of the mutation in pedigree samples.These results were consistent with those of high-throughput sequencing.CONCLUSIONS ANK2p.F3067L is considered a novel and potentially pathogenic gene mutation in DCM.
文摘Virion RNA was abstracted from purified type I Avian paramyxovirus strain YN-PA01(isolated from parrot)and used as a template. The fragment containing the fusion gene(F) and hemagglutinin-neuraminidase gene(HN) of the isolate was amplified by RT-PCR and cloned to the pMD18-T Vector. Using primer walking method the complete sequence of F-HN genes was obtained finally.And the respective amino acid sequence was deduced. Through relative software the phylogenetic trees on F gene and HN gene were constructed between strain YN-PA01 and reference strains. The results showed that strain YN-PA01 comparing with reference strains displays 98.7%-83.2% nucleotide homology and 98.1%-86.2% amino acid homology on F gene; 97.4%-79.1% nucleotide homology and 97.2%-83.2% amino acid homology on HN gene. Additional 6 amino acids are encoded by the HN gene ORF of strain YN-PA01 comparing with national reference strains.The studied strain YN-PA01 exhibits highest identities with strain JS/5[O1/Go either analyzed on F gene or HN gene.
