Chen Deng, Zhang Jun-wu
National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, CAMS and PUMC, Beijing 100005, China.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2005 Apr;27(2):194-8.
To clone and identify novel mouse aging-related genes.
The improved differential display reverse transcription-polymerase chain reaction method was used to analyze the differential expression of old and young BALB/C mouse cortex tissues. The novel expression sequence targets (EST) with differential expression were further confirmed by Northern blot and used as original sequence to clone the full-length cDNA sequence by bioinformatics technique. The reverse transcription products of BALB/C mouse cortex total RNA was used as the amplification template.
A total of 42 differentially expressed EST were obtained. Homologic analysis showed that 29 EST represented known cDNAs and 13 were probably new EST. One of the EST, which were confirmed to have down regulative expression in the old mouse cortex tissues by Northern blot analysis, was used to obtain the full-length cDNA sequences from reverse transcription products of total RNA of BALB/C mice cortex by bioinformation method. A 1 382 bp of cDNA sequence including the EST was obtained and designed as Arzc (GenBank accession number: AY344585). The Arzc cDNA contains an open reading frame of 261 bp and encodes a peptide of 86 amino acids residuals. FASTA analysis showed that the deduced peptide had a homology with a portion of integrase/recombinase, a member of a phage integrase family.
A novel mouse gene that is probably related to aging was identified and cloned.
克隆并鉴定新的小鼠衰老相关基因。
采用改进的差异显示逆转录-聚合酶链反应方法分析老年和幼年BALB/C小鼠皮质组织的差异表达。通过Northern印迹进一步确认差异表达的新表达序列标签(EST),并将其作为原始序列,利用生物信息学技术克隆全长cDNA序列。以BALB/C小鼠皮质总RNA的逆转录产物作为扩增模板。
共获得42个差异表达的EST。同源性分析表明,29个EST代表已知的cDNA,13个可能是新的EST。通过Northern印迹分析证实其中一个EST在老年小鼠皮质组织中表达下调,利用生物信息学方法从BALB/C小鼠皮质总RNA的逆转录产物中获得其全长cDNA序列。获得了一个包含该EST的1382 bp cDNA序列,并将其命名为Arzc(GenBank登录号:AY344585)。Arzc cDNA包含一个261 bp的开放阅读框,编码一个由86个氨基酸残基组成的肽段。FASTA分析表明,推导的肽段与噬菌体整合酶家族成员整合酶/重组酶的一部分具有同源性。
鉴定并克隆了一个可能与衰老相关的新的小鼠基因。