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Genotyping of bovine k‐casein (k‐CN A , k‐CN B , k‐CN C , k‐CN E ) following DNA sequence amplification and direct sequencing of k‐CN E PCR product
Author(s) -
SCHLIEBEN S.,
ERHARDT G.,
SENFT B.
Publication year - 1991
Publication title -
animal genetics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.756
H-Index - 81
eISSN - 1365-2052
pISSN - 0268-9146
DOI - 10.1111/j.1365-2052.1991.tb00687.x
Subject(s) - microbiology and biotechnology , biology , restriction enzyme , cytosine , polymerase chain reaction , genotyping , dna , genotype , gene , biochemistry
Summary. Genomic DNA isolated from blood and semen of dairy cattle with known k‐casein (k‐CN) genotypes was subjected to Southern blot hybridization and polymerase chain reaction (PCR) using up to 14 restriction endonucleases. k‐casein genotypes AA, AB and BB were identified using Hin dIII and Hin ft while genotypes with k‐CN C and k‐CN E were misidentified. Direct sequencing of the PCR product (k‐CN EE) showed a substitution of guanine (k‐CN A, B ) by adenine (k‐CN E ) which creates a Hae III restriction site. Therefore using PCR followed by Hin dIII or Hin fI and Hae III digest allows discrimination between k‐casein A, B and E directly at the DNA level.