Five different isoforms （IrlVHA-c1-c5） of V-ATPase subunit c （VHA-c） were cloned from a Japanese iris （Iris lactea Pall. var. chinensis Fisch. Koidz） cDNA library using degenerate primers PCR and the 5＇-RACE te...Five different isoforms （IrlVHA-c1-c5） of V-ATPase subunit c （VHA-c） were cloned from a Japanese iris （Iris lactea Pall. var. chinensis Fisch. Koidz） cDNA library using degenerate primers PCR and the 5＇-RACE technique. The sequence analysis showed the open reading frame （ORF） of the IrlVHA-c1 c5 to be 495 bp, corresponding to a protein of 164 amino acids. Among the five isoforms, IrlVHA-c1 and IrlVHA-c2 are completely homologous. The IrlVHA-c protein is localized at the vacuolar membrane as indicated by a green fluorescent protein （GFP） marker. Its over-expression in yeast could enhance yeast tolerance to NaCl stress. These results show that there are at least five genes encoding different isoforms of IrlVHA-c in Japanese iris and IrlVHA-c is important for the function of V-ATPase.展开更多
Vacuolar H^+-ATPase was regarded as a key enzyme promoting the fiber cell elongation in cotton (Gossypium hirsuturm L.) through regulating turgor-driven pressure involved in polarity expansion of single cell fiber. Th...Vacuolar H^+-ATPase was regarded as a key enzyme promoting the fiber cell elongation in cotton (Gossypium hirsuturm L.) through regulating turgor-driven pressure involved in polarity expansion of single cell fiber. The DET3, a V-ATPase subunit C, plays an important role in assembling subunits and regulating the enzyme activity, and is involved in Brassinosteroid-induced cell elongation. To analyze the function of GhDET3 on the elongation of cotton fibers, seven candidates of ESTs were screened and contigged for a 5'-upstream sequence, and the 3'-RACE technique was used to clone the 3'-downstream sequence for the full length of GhDET3 gene. The full length of the target clone was 1,340 bp, including a 10 bp 5'-UTR, an ORF of 1,134 bp, and a 196 bp 3'-UTR. This cDNA sequence encoded a polypepide of 377 amino acid residues with a predicted molecular mass of 43 kDa and a basic isoelectric point of 5.58. Furthermore, a length of 3,410 bp sequence from genomic DNA of GhDET3 was also cloned by PCR. The deduced amino acid sequence had a high homology with DET3 from Arabidopsis, rice, and maize. Quantitative real-time PCR (qRT-PCR) analysis showed that the GhDET3 expression pattern was ubiquitous in all the tissues and organs detected. The result also revealed that the accumulation of GhDET3 mRNA reached the highest profile at the fiber elongation stage in 12 DPA (days post anthesis) fibers, compared with the lowest level at the fiber initiation stage in 0 DPA ovules (with fibers). The transcript accumulation in fibers and ovules shared the similar variation tendency. In addition, in vitro ovule culture experiment demonstrated that exogenous 24-EBL treatment to 4 DPA ovules (with fibers) was capable of increasing the expression level of GhDET3, and the mRNA accumulation of GhDET3 increased in transgenic FBP7::GhDET2 cotton fibers in vivo. These results indicate that GhDET3 gene plays a crucial role in cotton fiber elongation.展开更多
目的:探讨膀胱癌细胞中细胞色素c氧化酶亚基7A2(COX7A2)基因的表达,及其与顺铂联用对膀胱癌J82细胞增殖、凋亡及线粒体功能影响。方法:采用生物信息学方法分析COX7A2在膀胱癌患者中的表达,并在J82细胞中进行验证。功能实验分为对照组(...目的:探讨膀胱癌细胞中细胞色素c氧化酶亚基7A2(COX7A2)基因的表达,及其与顺铂联用对膀胱癌J82细胞增殖、凋亡及线粒体功能影响。方法:采用生物信息学方法分析COX7A2在膀胱癌患者中的表达,并在J82细胞中进行验证。功能实验分为对照组(仅转染阴性对照siNC)、siRNA组(仅转染COX7A2的siRNA)、对照组+顺铂组(先转染阴性对照后用顺铂处理)和siRNA+顺铂组(先敲低COX7A2后用顺铂处理)。CCK-8、Transwell迁移能力测试和克隆增殖实验检测对照组和siRNA组中J82细胞的增殖、迁移能力。采用相应试剂盒检测各组细胞的ATP水平、活性氧(ROS)水平及线粒体膜电位(ΔΨm),以评估线粒体功能。流式细胞术检测各组细胞凋亡,以反映细胞的线粒体状态与对顺铂治疗的响应性关系。进一步通过癌症治疗响应基因标识数据库(CTR-DB),分析COX7A2与接受顺铂联合治疗的膀胱癌患者预后的关系。结果:生物信息学分析与生存曲线显示,COX7A2在膀胱癌患者中高表达并且与患者预后不良有关联。COX7A2在J82细胞中呈高表达(P<0.05)。在未经顺铂处理时,与对照组相比,siRNA组J82细胞增殖、迁移和克隆形成能力均显著下降(均P<0.001),而线粒体的ATP表达减少(P<0.01)、ROS表达量增多(P<0.0001)、MMP发生去极化(P<0.0001),凋亡水平增加(P<0.05);顺铂处理后,与对照组+顺铂相比,siRNA+顺铂组ATP表达减少(P<0.01)、ROS表达量增多(P<0.0001)、MMP发生去极化(P<0.0001),线粒体功能受损,凋亡水平增加(P<0.001)。CTR-DB数据库生信分析显示,5例接受顺铂+多柔比星+甲氨蝶呤+长春碱联合治疗的膀胱癌患者中,有应答者比无应答者COX7A2中位RNA表达量低(中位表达量:4501 vs 5009),12例铂类药物联合治疗的膀胱癌患者中有应答者比无应答者COX7A2中位RNA表达量低(中位表达量:2947 vs 3035),由于样本量有限,虽观察到趋势但无统计学意义。结论:敲低COX7A2可通过损伤线粒体功能,抑制膀胱癌细胞增殖与迁移,并可能由此增强细胞对顺铂诱导凋亡的敏感性。展开更多
Understanding the genetic diversity–area relationship(GAR)is essential for comprehending how species adapt to environmental changes,as genetic diversity is an indicator of a species’adaptive potential.Variation in e...Understanding the genetic diversity–area relationship(GAR)is essential for comprehending how species adapt to environmental changes,as genetic diversity is an indicator of a species’adaptive potential.Variation in environmental adaptation capacity exists among species and animal taxa with different distribution areas,highlighting the importance of understanding the GAR.To obtain a more comprehensive understanding of the GAR in terrestrial vertebrates,we assessed both haplotype diversity–area and nucleotide diversity–area relationships using 25,453 cytochrome c oxidase subunit I(COI)sequences from 142 amphibian species,574 bird species,and 342 mammal species.We found that both measures of genetic diversity increased with species range size across major animal groups.Nevertheless,the GAR did not differ among animal groups,while haplotype diversity performed better than nucleotide diversity in profiling the GAR,as indicated by higher R2 values.The difference in the modeling fit may stem from the distinct biological and mathematical significance of nucleotide diversity and haplotype diversity.These results suggest that the GAR follows similar rules among different animal taxa.Furthermore,haplotype diversity may serve as a more reliable indicator for assessing the potential effects of area size changes on animal populations and provide better guidance for conserving genetic diversity.展开更多
基金supported by the Heilongjiang Province Postdoctoral Science Foundation(LBH-Q10144)
文摘Five different isoforms （IrlVHA-c1-c5） of V-ATPase subunit c （VHA-c） were cloned from a Japanese iris （Iris lactea Pall. var. chinensis Fisch. Koidz） cDNA library using degenerate primers PCR and the 5＇-RACE technique. The sequence analysis showed the open reading frame （ORF） of the IrlVHA-c1 c5 to be 495 bp, corresponding to a protein of 164 amino acids. Among the five isoforms, IrlVHA-c1 and IrlVHA-c2 are completely homologous. The IrlVHA-c protein is localized at the vacuolar membrane as indicated by a green fluorescent protein （GFP） marker. Its over-expression in yeast could enhance yeast tolerance to NaCl stress. These results show that there are at least five genes encoding different isoforms of IrlVHA-c in Japanese iris and IrlVHA-c is important for the function of V-ATPase.
基金the National Natural Science Foundation of China (No. 30370904 , 30671258) the National High Technology Research and Development Program (863 Project) of China (No. 2006AA10Z121) the Program for New Century Excellent Talents in University (No. NCET-07-0712).
文摘Vacuolar H^+-ATPase was regarded as a key enzyme promoting the fiber cell elongation in cotton (Gossypium hirsuturm L.) through regulating turgor-driven pressure involved in polarity expansion of single cell fiber. The DET3, a V-ATPase subunit C, plays an important role in assembling subunits and regulating the enzyme activity, and is involved in Brassinosteroid-induced cell elongation. To analyze the function of GhDET3 on the elongation of cotton fibers, seven candidates of ESTs were screened and contigged for a 5'-upstream sequence, and the 3'-RACE technique was used to clone the 3'-downstream sequence for the full length of GhDET3 gene. The full length of the target clone was 1,340 bp, including a 10 bp 5'-UTR, an ORF of 1,134 bp, and a 196 bp 3'-UTR. This cDNA sequence encoded a polypepide of 377 amino acid residues with a predicted molecular mass of 43 kDa and a basic isoelectric point of 5.58. Furthermore, a length of 3,410 bp sequence from genomic DNA of GhDET3 was also cloned by PCR. The deduced amino acid sequence had a high homology with DET3 from Arabidopsis, rice, and maize. Quantitative real-time PCR (qRT-PCR) analysis showed that the GhDET3 expression pattern was ubiquitous in all the tissues and organs detected. The result also revealed that the accumulation of GhDET3 mRNA reached the highest profile at the fiber elongation stage in 12 DPA (days post anthesis) fibers, compared with the lowest level at the fiber initiation stage in 0 DPA ovules (with fibers). The transcript accumulation in fibers and ovules shared the similar variation tendency. In addition, in vitro ovule culture experiment demonstrated that exogenous 24-EBL treatment to 4 DPA ovules (with fibers) was capable of increasing the expression level of GhDET3, and the mRNA accumulation of GhDET3 increased in transgenic FBP7::GhDET2 cotton fibers in vivo. These results indicate that GhDET3 gene plays a crucial role in cotton fiber elongation.
文摘目的:探讨膀胱癌细胞中细胞色素c氧化酶亚基7A2(COX7A2)基因的表达,及其与顺铂联用对膀胱癌J82细胞增殖、凋亡及线粒体功能影响。方法:采用生物信息学方法分析COX7A2在膀胱癌患者中的表达,并在J82细胞中进行验证。功能实验分为对照组(仅转染阴性对照siNC)、siRNA组(仅转染COX7A2的siRNA)、对照组+顺铂组(先转染阴性对照后用顺铂处理)和siRNA+顺铂组(先敲低COX7A2后用顺铂处理)。CCK-8、Transwell迁移能力测试和克隆增殖实验检测对照组和siRNA组中J82细胞的增殖、迁移能力。采用相应试剂盒检测各组细胞的ATP水平、活性氧(ROS)水平及线粒体膜电位(ΔΨm),以评估线粒体功能。流式细胞术检测各组细胞凋亡,以反映细胞的线粒体状态与对顺铂治疗的响应性关系。进一步通过癌症治疗响应基因标识数据库(CTR-DB),分析COX7A2与接受顺铂联合治疗的膀胱癌患者预后的关系。结果:生物信息学分析与生存曲线显示,COX7A2在膀胱癌患者中高表达并且与患者预后不良有关联。COX7A2在J82细胞中呈高表达(P<0.05)。在未经顺铂处理时,与对照组相比,siRNA组J82细胞增殖、迁移和克隆形成能力均显著下降(均P<0.001),而线粒体的ATP表达减少(P<0.01)、ROS表达量增多(P<0.0001)、MMP发生去极化(P<0.0001),凋亡水平增加(P<0.05);顺铂处理后,与对照组+顺铂相比,siRNA+顺铂组ATP表达减少(P<0.01)、ROS表达量增多(P<0.0001)、MMP发生去极化(P<0.0001),线粒体功能受损,凋亡水平增加(P<0.001)。CTR-DB数据库生信分析显示,5例接受顺铂+多柔比星+甲氨蝶呤+长春碱联合治疗的膀胱癌患者中,有应答者比无应答者COX7A2中位RNA表达量低(中位表达量:4501 vs 5009),12例铂类药物联合治疗的膀胱癌患者中有应答者比无应答者COX7A2中位RNA表达量低(中位表达量:2947 vs 3035),由于样本量有限,虽观察到趋势但无统计学意义。结论:敲低COX7A2可通过损伤线粒体功能,抑制膀胱癌细胞增殖与迁移,并可能由此增强细胞对顺铂诱导凋亡的敏感性。
基金supported by the National Natural Science Foundation of China(32130013,32070434)the National Key Research and Development Program of China(2022YFC2601601)+1 种基金the Second Tibetan Plateau Scientific Expedition and Research(STEP)program(2019QZKK05010112,2019QZKK0304-02)Hainan Tropical Rainforest Conservation Research Project,ZDYF2023RDYL01(supported by the Hainan Institute of National Park,HINP,KY-24ZK02).
文摘Understanding the genetic diversity–area relationship(GAR)is essential for comprehending how species adapt to environmental changes,as genetic diversity is an indicator of a species’adaptive potential.Variation in environmental adaptation capacity exists among species and animal taxa with different distribution areas,highlighting the importance of understanding the GAR.To obtain a more comprehensive understanding of the GAR in terrestrial vertebrates,we assessed both haplotype diversity–area and nucleotide diversity–area relationships using 25,453 cytochrome c oxidase subunit I(COI)sequences from 142 amphibian species,574 bird species,and 342 mammal species.We found that both measures of genetic diversity increased with species range size across major animal groups.Nevertheless,the GAR did not differ among animal groups,while haplotype diversity performed better than nucleotide diversity in profiling the GAR,as indicated by higher R2 values.The difference in the modeling fit may stem from the distinct biological and mathematical significance of nucleotide diversity and haplotype diversity.These results suggest that the GAR follows similar rules among different animal taxa.Furthermore,haplotype diversity may serve as a more reliable indicator for assessing the potential effects of area size changes on animal populations and provide better guidance for conserving genetic diversity.
