Czech J. Anim. Sci., 2006, 51(10):425-428 | DOI: 10.17221/3960-CJAS
Cloning and sequencing of the complicated rDNA gene family of Bos taurus
- 1 College of Life Science, Foshan University, Foshan, Guangdong, P.R. China
- 2 College of Life Science and Technology, Jinan University, Guangzhou, Guangdong, P.R. China
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
Published: October 31, 2006 Show citation
References
- Chen S.W., Wang X.Z. (1996): The PCR methods of amplifying the large fragments of target DNA. Biotechnology, 6, 1-2.
- Gonzalez I.L., Sylvester J.E. (1995): Complete sequence of the 43-kb human ribosomal DNA repeat: analysis of the intergenic spacer. Genomics, 27, 320-328.
Go to original source...
Go to PubMed...
- He Z.Y., Li Y.Q. (2001): The role of dimethyl sulfoxide on polymerase chain reaction for human apoE gene. J. Jinan Univ. (Nat. Sci.), 22, 104-107.
- Li A.L., Jiang T., Ma Z.Y., Jia J.Z. (2002): Several methodologies on improving the PCR products. Biotechnol. Bull., 6, 33-35.
- Tang D.S., Shi J.Q., Tian Y.B., Gu W.J. (2002): Multiple site gene targeting in somatic cells and desired site integration of genes. J. Foshan Univ. (Nat. Sci. Ed.), 20, 64-68.
This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International (CC BY NC 4.0), which permits non-comercial use, distribution, and reproduction in any medium, provided the original publication is properly cited. No use, distribution or reproduction is permitted which does not comply with these terms.