TAR cloning 법에 의한 인간 및 마우스의 상동성 HPRT 유전자의 분리 = Isolation of Human and Mouse Orthologue HPRT Genes by Transformation-Associated Recombination (TAR) cloning

다국어 초록 (Multilingual Abstract)

The transformation-associated recombination (TAR) cloning technique allows selective isolation of chromosome regions or genes from complex genome. The procedure requires knowledge of relatively small genomic sequences that reside adjacent to the chromosome region of interest. This method involves homologous recombination during spheroplast transformation between genomic DNA and a TAR vector that has 5' and 3' gene targeting sequences (hooks). To examine whether TAR cloning can be applied to the isolation of gene homologues, we chose the HPRT genes from human and mouse genome. As results, the yield of positive clones for HPRT gene from human and mouse genome when using a TAR vector containing mHPRT hook or hHPRT hook was almost same level. Analysis of the gap regions in mHPRT revealed that they contain abnormalities that could result in instability of the sequences. In conclusion, we were able to use the TAR cloning technology to isolate gene homologue (orthologue) from nonidentical genome. Moreover, the use of the TAR cloning system may accelerate work on closing the remaining gaps in mammalian genome to achieve the goal of annotation of all mammalian genes.

참고문헌 (Reference)

1 "complete genomic sequence and analysis of tandem repeat polymorphisms in intronic regions" 21 : 769-777, 2002

2 "The utility of tar vectors used for selective gene isolation by TAR cloning" 31 : 322-328, 2003

3 "Substrate length requirements for efficient mitotic recombination in Saccharomyces cerevisiae" 3937-3950, 1993

4 "Structural instability of human tandemly repeated DNA sequences cloned in yeast artificial chromosome vectors" 1421-1428, 1990

5 "Specific isolation of human rDNA genes by TAR cloning" 197 : 269-276, 1997

6 "Specific cloning of human DNA as yeast artificial chromosomes by transformation associated recombination" 93 : 491-496, 1996

7 "Separation of long range human TERT gene haplotypes by transformation associated recombination cloning in yeast" 22 : 2452-2456, 2003

8 "Selective Isolation of mammalian Genes by TAR cloning In Current Protocols in Human Genetics" John Wiley Sons Inc 5-17, 1999

9 "Optimum conditions for selective isolation of genes from complex genomes by transformation associated recombination cloning" 31 : 29-, 2003

10 "Minimum length of sequence homology required for in vivo cloning by homologous recombination in yeast" 38 : 91-96, 1997

11 "Methods in yeast genetics" Cold Spring Harbor Laboratory Press 1986

12 "Isolation of a functional copy of the human BRCA1 gene by transformation associated recombination in yeast" 250 : 201-208, 2000

13 "Highly selective isolation of human DNAs from rodent human hybrid cells as circular yeast artificial chromosomes by transformation associated recombination cloning" 93 : 13925-13930, 1996

14 "Functional copies of a human gene can be directly isolated by transformation associated recombination cloning with a small 3 end target sequence" 95 : 4469-4474, 1998

15 "Effect of GC content on target hook required for gene isolation by transformation associated recombination cloning" 39 : 128-134, 2003

16 "Direct isolation of human BRCA2 gene by transformation associated recombination in yeast Proc" 94 : 7384-94 7387, 1997

17 "Direct cloning of human 10q25 neocentromere DNA using transformationassociated recombination" 47 : 399-404, 1997

18 "Defining the minimal length of sequence homology required for selective gene isolation by TAR cloning" 29 : 32-, 2002

19 "Closing the Gaps on Human Chromosome 19 Revealed Genes with a High Density of Repetitive Tandemly Arrayed Elements" 14 : 239-246, 2004

20 "Cloning of large segments of exogenous DNA into yeast by means of artificial chromosome vectors" 806-236 812, 1987

21 "A novel strategy for analysis of gene homologues and segmental genome duplications" 56 : 702-710, 2003