PMID- 27834335
OWN - NLM
STAT- MEDLINE
DCOM- 20170612
LR  - 20190320
IS  - 0971-5916 (Print)
IS  - 0971-5916 (Linking)
VI  - 144
IP  - 1
DP  - 2016 Jul
TI  - A PCR-RFLP method for detection of the LNPEP encoding human insulin-regulated 
      aminopeptidase (IRAP) rs4869317 polymorphism.
PG  - 120-123
LID - 10.4103/0971-5916.193298 [doi]
AB  - BACKGROUND & OBJECTIVES: Insulin regulated aminopeptidase (IRAP) has been related 
      to certain pathologies such as breast cancer, Alzheimer΄s disease and septic 
      shock. IRAP is encoded by the leucyl/cystinyl aminopeptidase (LNPEP) gene. The 
      genetic variation in the LNPEP gene has been analyzed in relation with the 
      mortality and vasopressin clearance in septic shock. The LNPEP rs4869317 SNP 
      (single nucleotide polymorphism) was the most significantly associated SNP with 
      vasopressinase activity, being TT genotype associated with increased mortality. 
      The objective of the present study was to develop a simple method to allow a 
      quick and affordable genotyping for the rs4869317 SNP of LNPEP gene. METHODS: 
      Blood DNA samples were obtained from randomly selected healthy volunteers (n=28). 
      A pair of primers was designed to amplify an 834 bp region of the LNPEP gene 
      containing the rs4869317 SNP. The two alleles (T or A) were detected by digestion 
      of the PCR products with the PacI restriction endonuclease. This enzyme only cuts 
      the PCR products when the adenine is present in the SNP. RESULTS: All individuals 
      showed RFPL (restriction fragment length polymorphism) fragments for the expected 
      genotypes (TT, TA or AA). The methodology was validated by sequencing of the 
      amplified DNAs from several 'T/T' and 'A/A' homozygotes and 'T/A' heterozygotes. 
      The results from both methods showed agreement. INTERPRETATION & CONCLUSIONS: The 
      PCR-RFLP is a simple and reliable method that allows a quick genotyping for the 
      rs4869317 SNP of LNPEP gene. The study of this polymorphism could be useful in 
      future investigations to analyze the role of genetic variants of IRAP in several 
      physiological/pathological conditions.
FAU - RamIrez-Exposito, MarIa Jesus
AU  - RamIrez-Exposito MJ
AD  - Experimental & Clinical Physiopathology Research Group, Department of Health 
      Sciences, University of Jaen, 23071 Jaen, Spain.
FAU - MartInez-Martos, Jos Manuel
AU  - MartInez-Martos JM
AD  - Experimental & Clinical Physiopathology Research Group, Department of Health 
      Sciences, University of Jaen, 23071 Jaen, Spain.
FAU - Palomeque, Teresa
AU  - Palomeque T
AD  - Unit of Genetics, Department of Experimental Biology, University of Jaen, 23071, 
      Jaen, Spain.
FAU - Lorite, Pedro
AU  - Lorite P
AD  - Unit of Genetics, Department of Experimental Biology, University of Jaen, 23071, 
      Jaen, Spain.
LA  - eng
PT  - Journal Article
PL  - India
TA  - Indian J Med Res
JT  - The Indian journal of medical research
JID - 0374701
RN  - 0 (DNA Primers)
RN  - EC 3.4.11.3 (Cystinyl Aminopeptidase)
RN  - EC 3.4.11.3 (leucyl-cystinyl aminopeptidase)
SB  - IM
MH  - Alleles
MH  - Cystinyl Aminopeptidase/genetics/*isolation & purification
MH  - DNA Primers
MH  - *Genotype
MH  - Humans
MH  - *Polymorphism, Restriction Fragment Length
MH  - Polymorphism, Single Nucleotide/genetics
MH  - Shock, Septic/*genetics/pathology
PMC - PMC5116884
COIS- None.
EDAT- 2016/11/12 06:00
MHDA- 2019/03/21 06:00
PMCR- 2016/07/01
CRDT- 2016/11/12 06:00
PHST- 2016/11/12 06:00 [entrez]
PHST- 2016/11/12 06:00 [pubmed]
PHST- 2019/03/21 06:00 [medline]
PHST- 2016/07/01 00:00 [pmc-release]
AID - IndianJMedRes_2016_144_1_120_193298 [pii]
AID - IJMR-144-120 [pii]
AID - 10.4103/0971-5916.193298 [doi]
PST - ppublish
SO  - Indian J Med Res. 2016 Jul;144(1):120-123. doi: 10.4103/0971-5916.193298.