At the early stage,the transcriptome sequencing technique was used to detect the differentially expressed gene CsFK111 between vine cucumber and dwarf cucumber D0462.The gene was cloned,and bioinformatics software too...At the early stage,the transcriptome sequencing technique was used to detect the differentially expressed gene CsFK111 between vine cucumber and dwarf cucumber D0462.The gene was cloned,and bioinformatics software tools were used to analyze and predict the gene family and this gene.There were 30 members of the cucumber F-box gene family.The coding region of the cucumber CsFK111 gene was full-length 1314 bp,which encoded 437 amino acids and was predicted to be located in the nucleus.The protein encoded by this gene was a non-transmembrane protein,and the prediction of the secondary structure showed thatβ-lamellar structure and irregular crimp were dominant.A comparison of the phylogenetic tree showed that it was closest to cantaloupe and belonged to the same branch.The results provided a basis for future study on the regulation mechanism of the CsFK111 gene on cucumber dwarfing and also laid a foundation for further study of FBK family proteins.展开更多
TDF1(transcription-drived fragment) was homologous to the predicted S. lycopersicum nonspecific lipid-transfer protein,nsLTP 2-like(91%), and it was significantly upregulated in response to C. fulvum(cladosporium fulv...TDF1(transcription-drived fragment) was homologous to the predicted S. lycopersicum nonspecific lipid-transfer protein,nsLTP 2-like(91%), and it was significantly upregulated in response to C. fulvum(cladosporium fulvum) infection in tomato plants.In this experiment, the full-length cDNA of nsLTP 2-like was cloned using RACE technology based on the sequence of TDF1(GenBank: JZ717725). A full-length, 625 bp(GenBank: KU366289), cDNA sequence, which with 98% similarity to nsLTP 2-like gene(GenBank: XM015233692) was obtained. This cDNA contains an ORF(open reading frame) with full-length of 345 bp, coding of 114 amino acids, including 12.3% Ala and Gly. Protein molecular weight was 11.51 ku, the isoelectric point(pI) was 8.99, and average overall hydrophilicity was 0.412, with one phosphorylation sites, belonging to volatile acidic nuclear protein. Secondary structure prediction showed that α-Helix accounts for 30.7%, extension chain for 12.28%, β-corner for 9.65%, and random coil for 47.37%. Through comparative analysis of the homology among species, it was found that the amino acid sequence of tomato nsLTP 2-like protein had a high similarity with other plants, and with a specific conserved sequence which might related features in nsLTP 2-like protein. It also be analyzed the gene expression pattern of tomato in different parts and under different stress conditions.The results showed that nsLTP 2-like gene was up-regulated in varying degrees, under the condition of cold stress, exogenous hormone spraying and cladosporium fulvum infection. Therefore, it was speculated that the gene played a role in response to abiotic and biotic stress in tomato.展开更多
克隆合作猪神经营养因子3基因(Neurotrophin3,NTF3)的编码区序列(Coding region sequence,CDS),预测分析其结构与功能,并检测基因在不同组织中的表达量,构建组织表达谱。通过PCR扩增、Sanger测序等方法获得合作猪NTF3基因CDS区序列,使...克隆合作猪神经营养因子3基因(Neurotrophin3,NTF3)的编码区序列(Coding region sequence,CDS),预测分析其结构与功能,并检测基因在不同组织中的表达量,构建组织表达谱。通过PCR扩增、Sanger测序等方法获得合作猪NTF3基因CDS区序列,使用生物信息学方法分析其结构,并使用RT-qPCR技术检测其在不同组织中的表达水平。结果显示,合作猪NTF3基因CDS区长774 bp,编码257个氨基酸;等电点(pI)为9.46,属于碱性蛋白;合作猪与猪、牛亲缘关系较近;二级结构主要为α-螺旋和无规则卷曲,三级结构主要由β-折叠、α-螺旋等组成;整条链中亲水性氨基酸残基数量多于疏水性氨基酸残基数量,是亲水性蛋白;无跨膜区域,是非跨膜蛋白;在37~58位氨基酸残基有一个线圈结构域、64~79位有一个低复杂度结构域和144~249位有一个神经生长因子结构域;存在信号肽,推测是分泌蛋白;NTF3基因在合作猪脾脏的表达量最高,回肠表达量最低。展开更多
基金Supported by the National Natural Science Foundation of China(32272724)the National Science Foundation of Heilongjiang Province,China(LH2019C033)。
文摘At the early stage,the transcriptome sequencing technique was used to detect the differentially expressed gene CsFK111 between vine cucumber and dwarf cucumber D0462.The gene was cloned,and bioinformatics software tools were used to analyze and predict the gene family and this gene.There were 30 members of the cucumber F-box gene family.The coding region of the cucumber CsFK111 gene was full-length 1314 bp,which encoded 437 amino acids and was predicted to be located in the nucleus.The protein encoded by this gene was a non-transmembrane protein,and the prediction of the secondary structure showed thatβ-lamellar structure and irregular crimp were dominant.A comparison of the phylogenetic tree showed that it was closest to cantaloupe and belonged to the same branch.The results provided a basis for future study on the regulation mechanism of the CsFK111 gene on cucumber dwarfing and also laid a foundation for further study of FBK family proteins.
基金Supported by the National Key R&D Program of China(2017YFD0101900)China Agriculture Research System(CARS-23-A-16)the Science Foundation of Heilongjiang Province(C2017024)
文摘TDF1(transcription-drived fragment) was homologous to the predicted S. lycopersicum nonspecific lipid-transfer protein,nsLTP 2-like(91%), and it was significantly upregulated in response to C. fulvum(cladosporium fulvum) infection in tomato plants.In this experiment, the full-length cDNA of nsLTP 2-like was cloned using RACE technology based on the sequence of TDF1(GenBank: JZ717725). A full-length, 625 bp(GenBank: KU366289), cDNA sequence, which with 98% similarity to nsLTP 2-like gene(GenBank: XM015233692) was obtained. This cDNA contains an ORF(open reading frame) with full-length of 345 bp, coding of 114 amino acids, including 12.3% Ala and Gly. Protein molecular weight was 11.51 ku, the isoelectric point(pI) was 8.99, and average overall hydrophilicity was 0.412, with one phosphorylation sites, belonging to volatile acidic nuclear protein. Secondary structure prediction showed that α-Helix accounts for 30.7%, extension chain for 12.28%, β-corner for 9.65%, and random coil for 47.37%. Through comparative analysis of the homology among species, it was found that the amino acid sequence of tomato nsLTP 2-like protein had a high similarity with other plants, and with a specific conserved sequence which might related features in nsLTP 2-like protein. It also be analyzed the gene expression pattern of tomato in different parts and under different stress conditions.The results showed that nsLTP 2-like gene was up-regulated in varying degrees, under the condition of cold stress, exogenous hormone spraying and cladosporium fulvum infection. Therefore, it was speculated that the gene played a role in response to abiotic and biotic stress in tomato.
文摘克隆合作猪神经营养因子3基因(Neurotrophin3,NTF3)的编码区序列(Coding region sequence,CDS),预测分析其结构与功能,并检测基因在不同组织中的表达量,构建组织表达谱。通过PCR扩增、Sanger测序等方法获得合作猪NTF3基因CDS区序列,使用生物信息学方法分析其结构,并使用RT-qPCR技术检测其在不同组织中的表达水平。结果显示,合作猪NTF3基因CDS区长774 bp,编码257个氨基酸;等电点(pI)为9.46,属于碱性蛋白;合作猪与猪、牛亲缘关系较近;二级结构主要为α-螺旋和无规则卷曲,三级结构主要由β-折叠、α-螺旋等组成;整条链中亲水性氨基酸残基数量多于疏水性氨基酸残基数量,是亲水性蛋白;无跨膜区域,是非跨膜蛋白;在37~58位氨基酸残基有一个线圈结构域、64~79位有一个低复杂度结构域和144~249位有一个神经生长因子结构域;存在信号肽,推测是分泌蛋白;NTF3基因在合作猪脾脏的表达量最高,回肠表达量最低。
