In this study, we isolated a WD40-repeat gene from Artemisia annua glandular trichomes. This gene shows 69.97% sequence similarity to Arabidopsis TTG1 at aminoacid level. Sub-cellular localization study shows that AaW...In this study, we isolated a WD40-repeat gene from Artemisia annua glandular trichomes. This gene shows 69.97% sequence similarity to Arabidopsis TTG1 at aminoacid level. Sub-cellular localization study shows that AaWD40 protein diffuses in both cell nucleus and cytosol. The correct nuclear localization of AaWD40 was observed when co-expressed with AabHLH, a putative A. thaliana AtTTG1 homologue cloned from Artemisia annua glandular trichomes. When AaWD40 gene was ectopically over expressed in Arabidopsis transparent testa glabrous1-1 (ttg1-1) mutants of A. thaliana, PAs production in seeds was restored, and the trichomeless phenotypes of mutant were rescued. Real-time PCR analysis results revealed that ETC1, CPC, TTG2 and BAN (the downstream targets of AtTTG1 depend on regulatory complex), which regulate the epidermal differentiation and anthocyanin biosynthesis were differentially expressed as a result of AaWD40 over expression. Furthermore, the CLV1, CLV2, CLV3 and WUS, which are required to maintain the stem-cell niche of Arabidopsis shoot apex, were also modulated by AaWD40 and Arabidopsis TTG1. The transcriptions of AP2/ERF, bHLH, MYB, WRKY and NACs family proteins, which are mostly involved in defense, stress response and development regulation, were remarkably modulated by AaWD40 over expression. We hypothesize that WD40 repeat proteins act as a crucial factor in regulating a wide variety of cellular functions in A. thaliana.展开更多
A pair of PCR primers was designed based on the conserved sequences of WD40 family gene of different varieties.The normolization library was screened by PCR methods,the cDNA insert size of positive clones were analyze...A pair of PCR primers was designed based on the conserved sequences of WD40 family gene of different varieties.The normolization library was screened by PCR methods,the cDNA insert size of positive clones were analyzed by PCR method.A full-length cDNA of WD40(GenBank accession number:EU219610)in cotton was obtained from sequencing.This gene is 1 796 bp in length,containing an open reading frame encoding 274 amino acids and a stop codon from 35th to 860th position.The bioinformatics characterization indicates that the protein is encoded by WD40 domain.pI and molecular weight of the protein encoded were predicted to be 4.24 and 29 kDa,respectively.The yielded gene was accondingly named as GDRP1.RT-PCR analysis showed that the expression of GDRP1 varied during the gland formation process.These results will be helpful for our further study on the gland formation of cotton.展开更多
目的分析和预测亚洲牛带绦虫WD40基因及其编码蛋白的结构和特性,用于指导其生物学功能的实验研究。方法利用美国国家生物技术信息中心(NCBI,http://www.ncbi.nlm.nih.gov/)、瑞士生物信息学研究所的蛋白分析专家系统(ExPASY,http://ca.e...目的分析和预测亚洲牛带绦虫WD40基因及其编码蛋白的结构和特性,用于指导其生物学功能的实验研究。方法利用美国国家生物技术信息中心(NCBI,http://www.ncbi.nlm.nih.gov/)、瑞士生物信息学研究所的蛋白分析专家系统(ExPASY,http://ca.expasy.org/)和IEDB Analysis Recource提供的各种生物信息学在线分析程序,结合其它生物信息学分析软件包如Pcgene和Vector NTI suite,从亚洲牛带绦虫全长cDNA质粒文库中识别WD40基因及其编码区,分析、预测该基因编码的蛋白质的理化特性、翻译后的修饰位点、功能域、亚细胞定位、拓扑结构、二级结构、三维空间构象等。结果该基因全长1208bp,编码区为81～1056bp,编码402个氨基酸,为全长基因;无跨膜区,具有多个磷酸化位点,蛋白的理化性质稳定,理论分子量为35942.4;没有质体、线粒体定位序列;预测该蛋白表面有三个亲水性强的线性表位和三个B细胞抗原表位。结论应用生物信息方法从亚洲牛带绦虫成虫cDNA文库中识别出了亚洲牛带绦虫WD40基因,并对其所编码的蛋白质的结构与功能进行了预测。展开更多
文摘In this study, we isolated a WD40-repeat gene from Artemisia annua glandular trichomes. This gene shows 69.97% sequence similarity to Arabidopsis TTG1 at aminoacid level. Sub-cellular localization study shows that AaWD40 protein diffuses in both cell nucleus and cytosol. The correct nuclear localization of AaWD40 was observed when co-expressed with AabHLH, a putative A. thaliana AtTTG1 homologue cloned from Artemisia annua glandular trichomes. When AaWD40 gene was ectopically over expressed in Arabidopsis transparent testa glabrous1-1 (ttg1-1) mutants of A. thaliana, PAs production in seeds was restored, and the trichomeless phenotypes of mutant were rescued. Real-time PCR analysis results revealed that ETC1, CPC, TTG2 and BAN (the downstream targets of AtTTG1 depend on regulatory complex), which regulate the epidermal differentiation and anthocyanin biosynthesis were differentially expressed as a result of AaWD40 over expression. Furthermore, the CLV1, CLV2, CLV3 and WUS, which are required to maintain the stem-cell niche of Arabidopsis shoot apex, were also modulated by AaWD40 and Arabidopsis TTG1. The transcriptions of AP2/ERF, bHLH, MYB, WRKY and NACs family proteins, which are mostly involved in defense, stress response and development regulation, were remarkably modulated by AaWD40 over expression. We hypothesize that WD40 repeat proteins act as a crucial factor in regulating a wide variety of cellular functions in A. thaliana.
基金Supported by National Natural Science Foundation of China(3044003230771311)Natural Science Foundation of Chongqing(cstc2007BB1328)~~
文摘A pair of PCR primers was designed based on the conserved sequences of WD40 family gene of different varieties.The normolization library was screened by PCR methods,the cDNA insert size of positive clones were analyzed by PCR method.A full-length cDNA of WD40(GenBank accession number:EU219610)in cotton was obtained from sequencing.This gene is 1 796 bp in length,containing an open reading frame encoding 274 amino acids and a stop codon from 35th to 860th position.The bioinformatics characterization indicates that the protein is encoded by WD40 domain.pI and molecular weight of the protein encoded were predicted to be 4.24 and 29 kDa,respectively.The yielded gene was accondingly named as GDRP1.RT-PCR analysis showed that the expression of GDRP1 varied during the gland formation process.These results will be helpful for our further study on the gland formation of cotton.
文摘目的分析和预测亚洲牛带绦虫WD40基因及其编码蛋白的结构和特性,用于指导其生物学功能的实验研究。方法利用美国国家生物技术信息中心(NCBI,http://www.ncbi.nlm.nih.gov/)、瑞士生物信息学研究所的蛋白分析专家系统(ExPASY,http://ca.expasy.org/)和IEDB Analysis Recource提供的各种生物信息学在线分析程序,结合其它生物信息学分析软件包如Pcgene和Vector NTI suite,从亚洲牛带绦虫全长cDNA质粒文库中识别WD40基因及其编码区,分析、预测该基因编码的蛋白质的理化特性、翻译后的修饰位点、功能域、亚细胞定位、拓扑结构、二级结构、三维空间构象等。结果该基因全长1208bp,编码区为81～1056bp,编码402个氨基酸,为全长基因;无跨膜区,具有多个磷酸化位点,蛋白的理化性质稳定,理论分子量为35942.4;没有质体、线粒体定位序列;预测该蛋白表面有三个亲水性强的线性表位和三个B细胞抗原表位。结论应用生物信息方法从亚洲牛带绦虫成虫cDNA文库中识别出了亚洲牛带绦虫WD40基因,并对其所编码的蛋白质的结构与功能进行了预测。
