Experiments were performed to evaluate the efficiency of inhibition of HSP70 gene expressionby antisense oligonucleotides complementary to the mRNA of HSP70 and to test the effects ofinhibition of HSP70 gene expressio...Experiments were performed to evaluate the efficiency of inhibition of HSP70 gene expressionby antisense oligonucleotides complementary to the mRNA of HSP70 and to test the effects ofinhibition of HSP70 gene expression on subsequent embryonic sensitivity to heat shock. Theresults showed that transfection of pre-implantation embryos at 4-cell stage with 5 Mantisense oligo had no effect on in vitro blastocyst development. However, transfection with10 to 40 M antisense oligo had reduced in vitro blastocyst development to 15, 10% and 0; Forthe embryos which exposed to 40 M As arrested at the 16-cell stage, there was no blastocystformation within the heat shock groups. In contrast, transfection had no effect on embryonicsensitivity to heat shock, above 25% of embryos developed to blastocyst stage in controlgroups.展开更多
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
A genomic hsp70 gene was isolated from a rice IR36 genomic library and 4 794 bp of the gene have been sequenoed. The 5’ flanking region of the gene contained a putative TATA box and a typical heat shock element seque...A genomic hsp70 gene was isolated from a rice IR36 genomic library and 4 794 bp of the gene have been sequenoed. The 5’ flanking region of the gene contained a putative TATA box and a typical heat shock element sequence 5’-CTcgGAAccTTCgAG-3’. The amino acid sequence of the rice HSP70 deduced from the coding region shared 84%-92% homologies with those of HSP70s from other plant species. An intron 1939bp long was identified in the coding region at the codon specifying amino acid 72 (Asp), the similar position introns occurring in other intron-containing hsp70 genes. In addition, another intron of 57 bp was found in the 3’-untranslated region in the rice hsp70 gene. Southern blot hybridization showed that rice hsp70 gene family contained at least three members. Analysis of the RNA leveis with the gene-specific and non-specific probes revealed that the rice hsp70 gene expressed at normal temperature and the expression was enhanced by heat shock treatment.展开更多
基金supported financially by National(30270957)Shandong(Y2003D03)Natural Science Foundation of China
文摘Experiments were performed to evaluate the efficiency of inhibition of HSP70 gene expressionby antisense oligonucleotides complementary to the mRNA of HSP70 and to test the effects ofinhibition of HSP70 gene expression on subsequent embryonic sensitivity to heat shock. Theresults showed that transfection of pre-implantation embryos at 4-cell stage with 5 Mantisense oligo had no effect on in vitro blastocyst development. However, transfection with10 to 40 M antisense oligo had reduced in vitro blastocyst development to 15, 10% and 0; Forthe embryos which exposed to 40 M As arrested at the 16-cell stage, there was no blastocystformation within the heat shock groups. In contrast, transfection had no effect on embryonicsensitivity to heat shock, above 25% of embryos developed to blastocyst stage in controlgroups.
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。
基金Project supported by the Rockefeller Foundation.
文摘A genomic hsp70 gene was isolated from a rice IR36 genomic library and 4 794 bp of the gene have been sequenoed. The 5’ flanking region of the gene contained a putative TATA box and a typical heat shock element sequence 5’-CTcgGAAccTTCgAG-3’. The amino acid sequence of the rice HSP70 deduced from the coding region shared 84%-92% homologies with those of HSP70s from other plant species. An intron 1939bp long was identified in the coding region at the codon specifying amino acid 72 (Asp), the similar position introns occurring in other intron-containing hsp70 genes. In addition, another intron of 57 bp was found in the 3’-untranslated region in the rice hsp70 gene. Southern blot hybridization showed that rice hsp70 gene family contained at least three members. Analysis of the RNA leveis with the gene-specific and non-specific probes revealed that the rice hsp70 gene expressed at normal temperature and the expression was enhanced by heat shock treatment.
