PML分子不同剪接体的分型及其应用  

Isolation Different PML Isoform by PCR Analysis

作　　者：沈宇清[1] 夏梅[1] 缪凤琴[1] 谢维[1] 张建琼[1] 

机构地区：[1]东南大学医学院遗传与发育生物学系,发育与疾病相关基因教育部重点实验室,江苏南京210009

出　　处：《南京师大学报（自然科学版）》2009年第3期88-91,98,共5页Journal of Nanjing Normal University(Natural Science Edition)

基　　金：国家自然科学基金(编号30371311)

摘　　要：早幼粒细胞白血病(Promyelocytic Leukemia,PML)基因位于15号染色体,含有9个外显子,通过对内含子不同的剪接方式产生7大类20余种不同的PML剪接体.为对PML不同剪接体进行分型,针对PML各剪接体C末端设计5对型特异性引物,通过PCR扩增产物的有无及大小来判断PML分子的型别.利用设计的引物,通过对已知类型的PML质粒进行分型,确定了分型引物的可行性.对一株肝癌细胞系QGY7701中表达的PML分子进行分型,发现其为PML剪接体IV型与II型的混合型.研究表明,利用所设计5对引物的扩增,可以判断出某种组织或细胞中表达的PML分子剪接体型别.PML （Promyelocytic Leukemia） gene is located on chromosome 15 and composed of nine exons. More than twenty different PML isoforms have been discovered and categorized into seven groups by different splicing pattern. In order to indentify different isoforms of the individual groups,five pairs of primer were designed to be used for isoform-specific PCR amplification according to the sequences at the C-terminal of PML gene. Analysis of the different isoforms was based on the presence and the size of the PCR product amplified by using all the five pairs of primers. Control experiment was done by analysis of a pc DNA3.1-GFP-PML plasmid which is known to be PMLIV. In addition,PML isoform in a liver cancer cell line QGY7701 was determined to be a mixture of PMLII and PMLIV by the same way. In conclusion,PML isoform of the seven groups can be classified by simply PCR amplification using the designed five pairs of primers.

关 键 词：PML基因 剪接体 PCR 

分 类 号：R735.7[医药卫生—肿瘤]