DNA SPECIFIC TO BOVINE MALE AND DISCRIMINATION OF BOVINE MALE AND FEMALE

摘要

PURPOSE:To discriminate bovine male and female simply and in high sensitivity by using bovine male specific DNA having a specific base sequence or a bovine female specific DNA hybridisable with the DNA as a primer and a probe. CONSTITUTION:Peripheral blood is collected from mature Holstein male and female, centrifuged separately, leukocyte fractions are treated with proteinase, RNase and phenol to extract DNAs, which are mixed with primers shown by formula I and formula II and thermostable polymerase, reacted at 60 deg.C for 2 minutes, at 80 deg.C for 2.5 minutes, further at 93 deg.C for 1.5 minute, amplified by repeating PCR operation 30 times, subjected to electrophoresis on agarose gel, a bond existing in male DNA but not in female DNA is selected and the base sequence is determined to give a bovine male specific DNA containing front part of IO-1 sequence shown by formula III or part thereof. The DNA is used as a probe to accurately discriminate bovine male and female.