热激蛋白70(heat shock protein 70,Hsp70)在植物发育过程以及响应生物和非生物胁迫中起着重要作用。为探究小麦Hsp70基因家族进化关系、功能以及表达模式,本研究对乌拉尔图小麦、拟斯卑尔脱山羊草、二粒小麦、粗山羊草以及普通小麦的Hs...热激蛋白70(heat shock protein 70,Hsp70)在植物发育过程以及响应生物和非生物胁迫中起着重要作用。为探究小麦Hsp70基因家族进化关系、功能以及表达模式,本研究对乌拉尔图小麦、拟斯卑尔脱山羊草、二粒小麦、粗山羊草以及普通小麦的Hsp70基因进行全面的生物信息学分析,并通过RT-qPCR方法分析其部分Hsp70基因在不同外源激素和环境胁迫条件下的表达模式。结果表明,从乌拉尔图小麦、拟斯卑尔脱山羊草、二粒小麦、粗山羊草和普通小麦5个物种中,分别鉴定出30、41、60、28和94个Hsp70基因;系统发育分析表明5个物种Hsp70家族成员分为5个亚家族组,每组成员数量不相等,其中大部分成员分布在第Ⅰ组,且同一亚家族中大多数的Hsp70成员具有相似的基因结构和保守基序;进一步综合分析5个物种Hsp70基因的染色体定位和重复事件,发现Hsp70基因在5个物种的各染色体上分布不均匀,此外从5个物种中共发现12个串联重复事件和110个片段复制事件,表明片段复制事件促进了小麦Hsp70基因家族的扩张;顺式作用元件分析表明,5个物种Hsp70基因的启动子区域存在多种光响应元件、逆境响应元件、激素响应元件以及生长发育调节元件;此外RT-qPCR结果表明,5个物种部分Hsp70基因在不同激素处理和逆境胁迫下具有不同程度的响应,在高温和干旱胁迫下,所选8个Hsp70基因均上调表达。小麦及其祖先物种Hsp70基因的鉴定及其进化过程为进一步研究Hsp70基因在小麦生长发育过程中的功能以及在逆境胁迫下的响应机制提供理论基础。展开更多
Degenerate primers are particularly useful in amplifying homologous genes from different organisms. This paper describes a method for designing degenerate primers for a given multiple alignment of DNA sequences of hsp...Degenerate primers are particularly useful in amplifying homologous genes from different organisms. This paper describes a method for designing degenerate primers for a given multiple alignment of DNA sequences of hsp70 gene family using ClustalW algorithm and detect the consensus region in gene family of hsp70 in a plant species which have not any recorded information about hsp70 gene family in the Genbank of National Centre of Biotechnology Information （NCBI） like Arando donax .The sequenced consensus sequence ofArando donax is considered a gene marker in building the genome map sequence. The BLASTn program is used to find a homology between more than one accession numbers of DNA sequences, （X67711.2） was for Oryza sativa （hsp70）, （AY372071. l） was for Nicotiana tabacum （hsp70） and （L41253.2） was for Lycopersicon esculentum （Hsc70）. In silco PCR module was performed to detect the melting temperatures （Tm） and predicted the PCR product size （783 bp）.The result of designed degenerate primers showed that there was a homology founded among the designed primers and the DNA templates of the recorded sequences （AY372071.1, X67711.2 and L41253.2） with at least 80% identity. The designed degenerate primers were used to isolate a consensus region ofhsp70 gene family ofArando donax at the expected molecular weight （783 bp）. The isolated PCR product, （783 bp） ofArando donax was sequenced and submitted to the Database of Japan （DDBJ） with accession number AB819871. The ORF finder tool translated the accession number AB819871 and gave a selected frame which used to build 3D structure model. In conclusion, this study focused on the importance of designing the degenerate primers to isolate the gene family and predict the 3D structure of gene family depending on the ORF finder tool of Genebank.展开更多
文摘热激蛋白70(heat shock protein 70,Hsp70)在植物发育过程以及响应生物和非生物胁迫中起着重要作用。为探究小麦Hsp70基因家族进化关系、功能以及表达模式,本研究对乌拉尔图小麦、拟斯卑尔脱山羊草、二粒小麦、粗山羊草以及普通小麦的Hsp70基因进行全面的生物信息学分析,并通过RT-qPCR方法分析其部分Hsp70基因在不同外源激素和环境胁迫条件下的表达模式。结果表明,从乌拉尔图小麦、拟斯卑尔脱山羊草、二粒小麦、粗山羊草和普通小麦5个物种中,分别鉴定出30、41、60、28和94个Hsp70基因;系统发育分析表明5个物种Hsp70家族成员分为5个亚家族组,每组成员数量不相等,其中大部分成员分布在第Ⅰ组,且同一亚家族中大多数的Hsp70成员具有相似的基因结构和保守基序;进一步综合分析5个物种Hsp70基因的染色体定位和重复事件,发现Hsp70基因在5个物种的各染色体上分布不均匀,此外从5个物种中共发现12个串联重复事件和110个片段复制事件,表明片段复制事件促进了小麦Hsp70基因家族的扩张;顺式作用元件分析表明,5个物种Hsp70基因的启动子区域存在多种光响应元件、逆境响应元件、激素响应元件以及生长发育调节元件;此外RT-qPCR结果表明,5个物种部分Hsp70基因在不同激素处理和逆境胁迫下具有不同程度的响应,在高温和干旱胁迫下,所选8个Hsp70基因均上调表达。小麦及其祖先物种Hsp70基因的鉴定及其进化过程为进一步研究Hsp70基因在小麦生长发育过程中的功能以及在逆境胁迫下的响应机制提供理论基础。
文摘Degenerate primers are particularly useful in amplifying homologous genes from different organisms. This paper describes a method for designing degenerate primers for a given multiple alignment of DNA sequences of hsp70 gene family using ClustalW algorithm and detect the consensus region in gene family of hsp70 in a plant species which have not any recorded information about hsp70 gene family in the Genbank of National Centre of Biotechnology Information （NCBI） like Arando donax .The sequenced consensus sequence ofArando donax is considered a gene marker in building the genome map sequence. The BLASTn program is used to find a homology between more than one accession numbers of DNA sequences, （X67711.2） was for Oryza sativa （hsp70）, （AY372071. l） was for Nicotiana tabacum （hsp70） and （L41253.2） was for Lycopersicon esculentum （Hsc70）. In silco PCR module was performed to detect the melting temperatures （Tm） and predicted the PCR product size （783 bp）.The result of designed degenerate primers showed that there was a homology founded among the designed primers and the DNA templates of the recorded sequences （AY372071.1, X67711.2 and L41253.2） with at least 80% identity. The designed degenerate primers were used to isolate a consensus region ofhsp70 gene family ofArando donax at the expected molecular weight （783 bp）. The isolated PCR product, （783 bp） ofArando donax was sequenced and submitted to the Database of Japan （DDBJ） with accession number AB819871. The ORF finder tool translated the accession number AB819871 and gave a selected frame which used to build 3D structure model. In conclusion, this study focused on the importance of designing the degenerate primers to isolate the gene family and predict the 3D structure of gene family depending on the ORF finder tool of Genebank.
