为了解60S核糖体蛋白L21(ribosomal protein L21,RPL21)的基因组结构和表达模式,以GenBank数据库中小麦RPL21基因的部分序列为信息探针,采用RT-PCR和PCR技术从小麦多子房株系幼穗中克隆了RPL21基因的cDNA与DNA片段,并对该基因在小麦多...为了解60S核糖体蛋白L21(ribosomal protein L21,RPL21)的基因组结构和表达模式,以GenBank数据库中小麦RPL21基因的部分序列为信息探针,采用RT-PCR和PCR技术从小麦多子房株系幼穗中克隆了RPL21基因的cDNA与DNA片段,并对该基因在小麦多子房近等基因系间的表达差异和多子房株系中的时空表达模式进行了分析。结果表明,所克隆的小麦RPL21基因cDNA序列(GenBank登录号:HM138480)长521bp,编码164个氨基酸;DNA序列(GenBank登录号:HM138481)长1600bp,含有2个外显子和1个内含子。半定量RT-PCR分析显示,该基因在多子房株系幼穗中表达少量上调;在多子房株系不同时期幼穗中的表达量随着幼穗的发育呈上升趋势;不同组织中具体表达模式为茎顶端≈幼根>幼穗>幼叶,生长旺盛部位表达量较高。展开更多
Ribosonal protein L21 which is a component of the 60s large ribosomal subunit plays an important role in ribosome. To explore the structure characteristic of ribosomal protein L21(rpL21) gene of the Ailuropoda melanol...Ribosonal protein L21 which is a component of the 60s large ribosomal subunit plays an important role in ribosome. To explore the structure characteristic of ribosomal protein L21(rpL21) gene of the Ailuropoda melanoleuca and investigate its homologies with other already reported sequenses' including Rattus norvegicus, Mus musculus, Mus musculus, etc. The cDNA of rpL21 was cloned from the Ailuropoda melanoleuca by RT-PCR. The sequence data were analyzed by Genscan software. Blast 2.1 was used to study the homology of the obtained rpL21 sequence with the gene sequence of other species; Open reading frame (ORF) of the DNA sequence was searched using ORF finder software; Protein structure of the rpL21 sequence cloned was deduced using Predict Protein software. The results indicated that the length of cDNA fragment cloned was 504bp; containing an open reading frame of 483bp. Deduced protein was composed of 160 amino acids with an estimated molecular weight of 18.59kD and pI of 11.10. The length of the genomic sequence is 2254bp, containing 5 exons and 4 introns. Alignment analysis indicates that rpL21 is highly similarity with the reported species both at the level of DNA and protein. Topology prediction shows that 6 different patterns were found in the rpL21 protein of the Ailuropoda melanoleuca. The rpL21 gene of the Ailuropoda melanoleuca was studied in this paper, which will enrich and improve the mammals' rpL21 gene database.展开更多
文摘为了解60S核糖体蛋白L21(ribosomal protein L21,RPL21)的基因组结构和表达模式,以GenBank数据库中小麦RPL21基因的部分序列为信息探针,采用RT-PCR和PCR技术从小麦多子房株系幼穗中克隆了RPL21基因的cDNA与DNA片段,并对该基因在小麦多子房近等基因系间的表达差异和多子房株系中的时空表达模式进行了分析。结果表明,所克隆的小麦RPL21基因cDNA序列(GenBank登录号:HM138480)长521bp,编码164个氨基酸;DNA序列(GenBank登录号:HM138481)长1600bp,含有2个外显子和1个内含子。半定量RT-PCR分析显示,该基因在多子房株系幼穗中表达少量上调;在多子房株系不同时期幼穗中的表达量随着幼穗的发育呈上升趋势;不同组织中具体表达模式为茎顶端≈幼根>幼穗>幼叶,生长旺盛部位表达量较高。
基金financially supported by Application Foundation Project in Sichuan Province(2009JY0061)Youth Fund Project of Educational Committee of Sichuan province(09ZB088)
文摘Ribosonal protein L21 which is a component of the 60s large ribosomal subunit plays an important role in ribosome. To explore the structure characteristic of ribosomal protein L21(rpL21) gene of the Ailuropoda melanoleuca and investigate its homologies with other already reported sequenses' including Rattus norvegicus, Mus musculus, Mus musculus, etc. The cDNA of rpL21 was cloned from the Ailuropoda melanoleuca by RT-PCR. The sequence data were analyzed by Genscan software. Blast 2.1 was used to study the homology of the obtained rpL21 sequence with the gene sequence of other species; Open reading frame (ORF) of the DNA sequence was searched using ORF finder software; Protein structure of the rpL21 sequence cloned was deduced using Predict Protein software. The results indicated that the length of cDNA fragment cloned was 504bp; containing an open reading frame of 483bp. Deduced protein was composed of 160 amino acids with an estimated molecular weight of 18.59kD and pI of 11.10. The length of the genomic sequence is 2254bp, containing 5 exons and 4 introns. Alignment analysis indicates that rpL21 is highly similarity with the reported species both at the level of DNA and protein. Topology prediction shows that 6 different patterns were found in the rpL21 protein of the Ailuropoda melanoleuca. The rpL21 gene of the Ailuropoda melanoleuca was studied in this paper, which will enrich and improve the mammals' rpL21 gene database.
