Cerit, H. “Determination of the Allele Frequency of Some Genomic Loci in a Holstein Cattle Population and its Importance in Individual Identification”


CERİT H.

Turk. J. Vet. Anim. Sci., cilt.27, ss.81-91, 2003 (SCI Expanded İndekslerine Giren Dergi)

  • Cilt numarası: 27
  • Basım Tarihi: 2003
  • Dergi Adı: Turk. J. Vet. Anim. Sci.
  • Sayfa Sayıları: ss.81-91

Özet

Son yÝllarda DNA dizilerinin deÛißik molek?ler genetik teknikleri ve ?zg?n DNA polimorfizmlerinin genetik marker olarak
kullanÝmlarÝ hÝzla gelißmektedir. Bu alÝßmada sÝÛÝr genomunda bulunan 7 deÛißik polimorfik lokusun allel ve genotip frekanslarÝnÝn
Üstanbul ?niversitesi Veteriner Fak?ltesi EÛitim, ?Ûretim, AraßtÝrma ve Uygulama ?iftliÛiÕndeki sÝÛÝr populasyonuna ait daÛÝlÝmlarÝ
incelenmißtir. Buna baÛlÝ olarak alÝßmanÝn hedefi araßtÝrmada ele alÝnan 7 lokusun heterozigotluk oranÝ dÝßlama g?c?, uyußma
olasÝlÝÛÝ ve ayrÝmlama g?c? aÝsÝndan deÛerlendirilmesi ve ilgili lokuslarÝn hayvanlarÝn kimlik tayininde kullanÝlabilirliklerinin
araßtÝrÝlmasÝ olmußtur.
Hayvanlardan alÝnan kan ?rneklerinden fenol-kloroform-izoamil alkol ve chelex y?ntemi kullanÝlarak DNA ekstraksiyonu yapÝlmÝßtÝr.
Spektrofotometre ile, DNAÕnÝn miktarÝ ve saflÝk derecesi belirlendikten sonra 7 DNA lokusu iin PCR y?ntemine dayalÝ amplifikasyon
yapÝlmÝßtÝr. Amplifikasyon ?r?nlerinin varlÝÛÝ EtBr ile boyanarak % 1,5 agaroz jel elektroforezi ile incelenmißtir. Denat?re edilmiß
poliakrilamid jel elektroforezi kullanÝlarak ve g?m?ß boyama yapÝlarak PCR ?r?nleri deÛerlendirilerek allellerin fenotipleri
belirlenmißtir.
AraßtÝrma sonunda BMS1822 lokusu dÝßÝnda Hardy-Weinberg dengesine uyumsuzluk g?zlenmemißtir. Üstanbul ?niversitesi Veteriner
Fak?ltesi EÛitim, ?Ûretim, AraßtÝrma ve Uygulama ?iftliÛi genelinde yapÝlan bu alÝßmada 7 STR/mikrosatellit lokusu iinde en y?ksek
heterozigotluk deÛeri % 83,3 ile BMS2270 lokusunda g?zlenirken en d?ß?k deÛer % 31,4 ile BMS2721 lokusunda g?r?lm?ßt?r.
En y?ksek dÝßlama olasÝlÝÛÝ 0,662 ile BMS2270 lokusunda g?zlenirken en d?ß?k deÛer 0,066 ile BMS2721 lokusunda g?zlenmißtir.
En y?ksek ayrÝmlama g?c? 0,947 ile BMS2270 lokusunda g?zlenirken en d?ß?k deÛer 0,504 ile BMS2721 lokusunda
g?zlemlenmißtir. Uyußma (karßÝlaßma) olasÝlÝÛÝnda ise en y?ksek deÛeri veren lokuslar BMS3019 ve BMS2721 olurken en d?ß?k
deÛerler BMS2270 ile BMS1822 lokuslarÝnda saptanmÝßtÝr. Bu alÝßmada kullanÝlan lokuslarÝn kombine ayrÝmlama ve dÝßlama g?c?
deÛerlerinin olduka y?ksek olduÛu g?zlenmißtir. DolayÝsÝyla bu sistemlerin DNA profilleme analizlerinde kullanÝlabilecek markerler
olabileceÛi s?ylenebilir.
Nowadays, the use of DNA sequences with genetic techniques and DNA polymorphism as genetic markers is increasing
very rapidly. In this study, we examined the frequencies of seven different polymorphic loci in the cattle population of Üstanbul
University Veterinary Medicine Research and Application Farm. Thus, our aim was to examine and calculate the power of exclusion,
match probability and power of discrimination of the seven loci, and examine the usefulness of the involved loci in identification tests.
For DNA extraction, Chelex and Phenol-Chloroform-Isoamil alcohol methods were applied to the blood taken from animals. After
measuring the amount and purity degree of DNA with a spectrophotometer, seven DNA loci were amplified by PCR. The amplification
products were stained with EtBr and examined with 1.5% Agarose gel electrophoresis. Using denaturated polyacrylamide gel
electrophoresis and silver staining, PCR products were evaluated and phenotypes of the alleles were determined.
At the end of the study, a mismatch to the Hardy-Weinberg equilibrium was observed in all loci but BMS1822. The highest
heterozygosity ratio was 83.3% for BMS2270, and the lowest ratio was 31.4% for BMS2721 among the seven STR/microsatellite
loci we studied. The highest exclusion probability was 0.662 for BMS 2270, and the lowest was 0.066 for BMS2721. The highest
power of discrimination was 0.947 for BMS2270, and the lowest was 0.504 for BMS2721. The highest matching probability was
for BMS3019 and BMS2721, and the lowest was for BMS2270 and BMS1822. It was observed that the loci in this study have very
high power of discrimination and power of exclusion values when combined. Thus, these systems may be useful as markers in DNA
profiling.