Cloning and sequencing of the complicated rDNA gene family of Bos taurus
D. Tang, H. Jiang, Y. Zhang, Y. Li, X. Zhang, T. Zhouhttps://doi.org/10.17221/3960-CJASCitation:Tang D., Jiang H., Zhang Y., Li Y., Zhang X., Zhou T. (2006): Cloning and sequencing of the complicated rDNA gene family of Bos taurus. Czech J. Anim. Sci., 51: 425-428.
The rDNA genes coding for ribosomal RNA (rRNA) in animals are repeat sequences with high GC content and complicated structure. Based on the sequences of human ribosomal DNA repeat unit and transcription unit and the long and accurate PCR method with LA Taq DNA polymerase and GC buffer, we were able to amplify the complicated repeat sequences of rDNA genes in Bos taurus. Three rDNA genes and 2 internal transcribed spacer (ITS) fragments were cloned and confirmed by sequencing. The conditions for the cloning of complicated DNA sequences such as special rules of primer design, improvement of the reaction system, selection of DNA polymerase and adjustment of cycle parameters were discussed.Keywords:
GC-rich; repetitive sequences; PCR; rDNA; rRNA; Bos Taurus