5′和3′末端快速扩增法克隆多发性骨髓瘤负性相关基因DAZAP2

目的在多发性骨髓瘤患者中KIAA0058基因序列表达明显下调,从正常人骨髓单个核细胞中克隆该序列全长cDNA,以期更好地研究该序列结构,获得其功能信息。方法采用末端快速扩增法(RACE),进行多轮RT-PCR后,对PCR产物进行测序、拼接、证实,与NCBI核酸数据库进行比对。结果该序列全长cDNA长度为1913bp,登陆号为AY430097,与来源于人睾丸cDNA文库的序列DAZAP2同源性高达99%以上。该序列具有较短的5′非翻译区及长的3′非翻译区,开放阅读框序列从第84位碱基至590位碱基,具有poly(A)尾。结论利用RACE法,从骨髓单个核细胞中成功克隆了DAZAP2全长cDNA。

Cloning of a multiple myeloma negative-related gene DAZAP2 by 5′ and 3′ rapid amplification of cDNA ends

ObjectiveTo get the full cDNA sequence of KIAA0058 from bone marrow mononuclear cells for better studying it's structure and function. MethodsThe KIAA0058 sequence was cloned by rapid amplification of cDNA ends(RACE) and pieced together these sequences. Then verified it by sequencing and BLASTN with NCBI nucleotide data bank. ResultsA 1913 bp sequence was obtained and deposited to Genebank (AY430097), which share high homolog (99%) to DAZAP2 from human testis cDNA library. The DAZAP2 gene contains a short 5′ UTR (untranslated region), an ORF (open reading frame) from nt84 to 590, and a relatively long 3′ UTR plus a poly(A) tail. ConclusionWe succeed to clone the full cDNA sequence of DAZAP2 from bone marrow mononuclear cells by RACE.