家族性急性髓系白血病相关新基因FAMLF cDNA全长的克隆及其生物功能分析  

Full-length cDNA cloning and biological function analysis of a novel gene FAMLF related to familial acute myelogenous leukemia

作　　者：李景岗[1] 王少元[1] 黄源茂[1] 王程毅[1] 

机构地区：[1]福建医科大学附属协和医院血液科福建省血液病研究所,福州350001

出　　处：《中华医学杂志》2008年第38期2667-2671,共5页National Medical Journal of China

基　　金：国家自然科学基金资助项目（30770909）;福建省重大科研项目基金资助（2003F003）;志谢 福建医科大学分子医学研究中心林旭教授提供技术指导

摘　　要：目的克隆家族性急性髓系白血病（AML）相关新基因cDNA全长，在分子水平上探讨急性白血病发生发展的机制。方法以构建的家族性AML抑制性消减文库中1个有差异表达的新基因EST序列zywb87（GenBank注册号CV973101）为基础，应用cDNA末端快速扩增法（RACE）克隆其全长cDNA，应用生物信息学对其功能进行初步分析，应用一步法半定量RT—PCR检测其在AML患者及正常人中的表达。结果获得了家族性AML相关新基因FAMLF的cDNA全长，该基因定位于染色体1q31．3，cDNA全长2313bp，开放读码框（ORF）为249bp，编码82个氨基酸的蛋白质，含有信号肽，富含亮氨酸重复单位（LRR—SD22），内在固有无序结构域等功能区。Blast检索为功能未知的新基因，已被GenBank收录，并命名为FAMLF，核酸注册号为EF413001，蛋白质注册号为ABN58747。新基因FAMLF在AML患者中的表达明显高于正常人，差异有统计学意义（2．61±0．66 vs 0．97±0．51，P〈0．01）。结论成功获得一个家族性AML相关新基因FAMLF cDNA全长，FAMLF基因在AML中高表达，可能是具有一定生物功能的新基因。Objective To clone the full-length cDNA of a novel gene related to familial acute myelogenous leukemia （AML） and to demonstrate its molecular mechanisms on the gene level. Methods Bone marrow specimen was obtained from a patient of familial AML, male, aged 11, and peripheral blood samples were obtained from 23 AML patients outside this family, 9 normal persons in this family, and 23 normal persons outside this family. Based on the EST sequence zywb87 （GenBank accession number： CV973101 ） from a subtractive cDNA library of differential expressed genes constructed in familial AML, SMART-rapid amplification of cDNA ends （ SMART-RACE ） was applied to clone the full-length cDNA of the novel gene, and bioinformatics was used to predict its biological function, the expression of the novel gene in AML was detected by One-Step RT-PCR. Results A full-length cDNA of 2313 bp was obtained from the bone marrow specimen of the familial AML patient with complete open reading frame （ORF） of 249 bp. Localized on 1 q31. 3 of human chromosome, it coded a 82-amino acid polypeptide with signal peptide, leucine-rieh repeat （LRR_SD22）, and intrinsic disorder functional domain. BLAST analysis conffirmed this gene as a novel gene designated with the accession number： （nucleotide） EF413001 and （protein） ABN58747 by GenBank and was named as Homo sapiens familial acute myelogenous leukemia related factor（ FAMLF）. The FAMLF expression level of the AML patients outside this family was （2.61 ± 0.66 ）, significantly higher than that of the normal persons outside this family （0.97 ± 0.51, P 〈 0.01 ）. Conclusion A full- length cDNA of the novel gene FAMLF related to familial AML has been obtained. The FAMLF gene is expressed highly in AML and may present biological function on the progress of AML.

关 键 词：白血病 髓样 基因 克隆 生物 计算生物学 cDNA末端快速扩增法 

分 类 号：R686[医药卫生—骨科学]