TITLE:
The Analysis of Primer Gene of Phosphodiesterase Type 5 (PDE5) on Erectyle Dysfungction
AUTHORS:
Theopilus W. Watuguly, Indranila K. Samsuria, Pudji Astuti, Tri Rini Nuringtyas, Nastiti Wijayanti, Syahran Wael
KEYWORDS:
Erectile Disfunction, PDE5, Primer, PCR
JOURNAL NAME:
Open Journal of Applied Sciences,
Vol.8 No.9,
September
28,
2018
ABSTRACT: Backround: Erectile dysfunction (ED) is the inability of male reproductive organs
within sexual intercourse caused by neurogenic and hormonal disorders. There
are some causes of ED such as hypertension, stress, neurological disorders,
stroke, diabetes, atherosclerosis, lifestyle, alcohol, smoking, and age-related
hormonal decline can cause infertility. The natural treatment of sexual
dysfunction through aphrodisiac activity of the plant is to increase sexual
hormones, spermatogenesis activity and through PDE5 inhibitors such as
sildenafil, verdenafil, and tadalafil which can inhibit the hydrolysis of second
messenger cGMP of penis smooth muscle cells. Primer
Design for amplification of several PDE5 gene nucleotide sequences obtained
from NCBI gen banks and tested directly through explosions at NCBI and also
using MEGA 6, primer 3 plus, and fast PCR software. The natural treatment of sexual dysfunction through aphrodisiac activity from plants to increase sexual hormone,
spermatogenesis activity and inhibitor PDE5 such as sildenafil, vardenafil, and
tadalafil can inhibit hydrolysis second messenger of cells cGMP smooth penis
muscles. Method: Primer design stages for several sequences are data
supply, multiple sequence alignment, sequence trimming, primer design (fastPCR
input), in silico PCR analysis, and primer evaluation (Primer Test, OligoCalc
and BLAST). Results: Primer of PDE5 that is choosen is with reverse sequence
5-TGCATTGACCATGTCTCTCGTT-3, forward
5-CGCCGATCTGGGCTGAACTA-3 able to amplify template DNA at temperature 67.2°C, 65.8°C,
63.7°C, however, the DNA band fragment looks not very clear, while it is more
clearly seen at Tm temperature 61.2°C, 59.1°C, 57.8°C and 57°C. Conclusion: PDE5 primers can be amplified well at temperature 61.2°C, 59.1°C, 57.8°C and 57°C. PDE5 primer succeeded in amplifying DNA
with a product length of 402 bp.