Duan Xiaoran, Li Chunyang, Wang Tuanwei, Feng Xiaolei, Cui Liuxin, Yao Wu, Wang Wei
Clin Lab. 2016;62(6):1179-82. doi: 10.7754/clin.lab.2015.150942.
In recent research, it has been shown that there have been variants of rs894160 within the PLINI gene which have been associated with obesity, type 2 diabetes, and other diseases. But the isoschizomers such as the Mn1I enzyme required for the detection of this polymorphism are expensive.
The study used an improved PCR-RFLP method with mismatched base for detection of the single nucleotide polymorphism rs894160.
After detecting 550 Chinese Han individuals, the genotype frequencies were 26.0% for AA, 50.0% for AG, and 24.0% for GG. The allelic frequencies were 51.0% for A and 49.0% for G. The PCR results were confirmed by DNA sequencing. The chi2 test showed the genotype and allele frequencies of PLIN1 do not deviate from Hardy-Weinberg equilibrium, and the sequences of amplified products were consistent with the one published in Genbank with the exception of mismatched base.
Based on the PCR with mismatched primers we designed, the PLIN1 polymorphisms could be identified effectively.
近期研究表明，PLIN1基因内的rs894160存在多种变体，这些变体与肥胖症、2型糖尿病及其他疾病有关。但检测该多态性所需的同裂酶，如Mn1I酶，价格昂贵。
本研究采用改进的带有错配碱基的PCR-RFLP方法检测单核苷酸多态性rs894160。
对550名中国汉族个体进行检测后，AA基因型频率为26.0%，AG基因型频率为50.0%，GG基因型频率为24.0%。等位基因频率A为51.0%，G为49.0%。PCR结果经DNA测序确认。卡方检验显示，PLIN1的基因型和等位基因频率未偏离哈迪-温伯格平衡，除错配碱基外，扩增产物序列与Genbank中公布的序列一致。
基于我们设计的错配引物PCR，可以有效鉴定PLIN1多态性。
