Functional cloning
The functional cloning includes methods for identification of unknown variants of a known gene , for example. B. from other species or different isoforms or homologous genes within an organism. In contrast to positional cloning , functional cloning is based on the search for a gene with a similar DNA sequence and therefore presumably a similar function.
properties
The DNA is purified by DNA isolation and then fragmented . By agarose gel electrophoresis , the DNA is fractionated and by Southern blot transferred to a membrane. By hybridization with a hybridization probe whose DNA sequence is based on the known gene, similar sequences are bound on the membrane. The sequences of the binding DNA fragments are determined by DNA sequencing .
Individual evidence
- ^ Y. Murakami: Functional cloning of a tumor suppressor gene, TSLC1, in human non-small cell lung cancer. In: Oncogene. Volume 21, Number 45, October 2002, pp. 6936-6948, ISSN 0950-9232 . doi : 10.1038 / sj.onc.1205825 . PMID 12362275 . PDF .
- ↑ R. Visconti, L. D'Adamio: Functional cloning of genes regulating apoptosis in neuronal cells. In: Methods in molecular biology (Clifton, NJ). Volume 399, 2007, pp. 125-131, ISSN 1064-3745 . doi : 10.1007 / 978-1-59745-504-6_9 . PMID 18309929 .
- ↑ JL Jestin: Functional cloning by phage display. In: Biochemistry. Volume 90, Number 9, September 2008, pp. 1273-1278, ISSN 0300-9084 . doi : 10.1016 / j.biochi.2008.04.003 . PMID 18466773 .