An Improved PCR-RFLP Assay for the Detection of a Polymorphism of PLIN1 Gene

Clin Lab. 2016;62(6):1179-82. doi: 10.7754/clin.lab.2015.150942.

Abstract

Background: 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.

Methods: The study used an improved PCR-RFLP method with mismatched base for detection of the single nucleotide polymorphism rs894160.

Results: 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.

Conclusions: Based on the PCR with mismatched primers we designed, the PLIN1 polymorphisms could be identified effectively.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amplified Fragment Length Polymorphism Analysis*
  • Asian People / genetics
  • Carrier Proteins / genetics*
  • Chi-Square Distribution
  • China
  • DNA Primers
  • Gene Frequency
  • Genotype
  • Humans
  • Perilipin-1
  • Phosphoproteins / genetics*
  • Polymorphism, Restriction Fragment Length*
  • Polymorphism, Single Nucleotide*
  • Predictive Value of Tests

Substances

  • Carrier Proteins
  • DNA Primers
  • PLIN1 protein, human
  • Perilipin-1
  • Phosphoproteins