PMID: 9539761May 16, 1998Paper

Functional copies of a human gene can be directly isolated by transformation-associated recombination cloning with a small 3' end target sequence

Proceedings of the National Academy of Sciences of the United States of America
Natalay KouprinaVladimir Larionov

Abstract

Unique, small sequences (sequence tag sites) have been identified at the 3' ends of most human genes that serve as landmarks in genome mapping. We investigated whether a single copy gene could be isolated directly from total human DNA by transformation-associated recombination (TAR) cloning in yeast using a short, 3' unique target. A TAR cloning vector was constructed that, when linearized, contained a small amount (381 bp) of 3' hypoxanthine phosphoribosyltransferase (HPRT) sequence at one end and an 189-bp Alu repeat at the other end. Transformation with this vector along with human DNA led to selective isolations of the entire HPRT gene as yeast artificial chromosomes (YACs) that extended from the 3' end sequence to various Alu positions as much as 600 kb upstream. These YACs were retrofitted with a NeoR and a bacterial artificial chromosome (BAC) sequence to transfer the YACs to bacteria and subsequently the BACs to mouse cells by using a Neo selection. Most of the HPRT isolates were functional, demonstrating that TAR cloning retains the functional integrity of the isolated material. Thus, this modified version of TAR cloning, which we refer to as radial TAR cloning, can be used to isolate large segments of the human genome...Continue Reading

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Citations

Jul 9, 2011·Pharmaceutical Research·Brian W BiggerCharles Coutelle
Sep 20, 2006·Nature Reviews. Genetics·Natalay Kouprina, Vladimir Larionov
Jul 19, 2000·The EMBO Journal·K S LobachevM A Resnick
Jan 15, 2002·Nucleic Acids Research·Vladimir NoskovVladimir Larionov
Aug 19, 2007·The Plant Cell·Andreas Houben, Ingo Schubert
Feb 22, 2012·Genome Biology·Yvonne Y ChenChristina D Smolke
Mar 22, 2001·Pharmacogenomics·R JudsonA Windemuth
Mar 22, 2001·Pharmacogenomics·R Judson, J C Stephens
Apr 6, 2013·Nucleic Acids Research·Artem V KononenkoVladimir Larionov
Oct 23, 2004·Genomics·Chang-En YuGerard D Schellenberg
Nov 26, 2003·FEMS Microbiology Reviews·Natalay Kouprina, Vladimir Larionov
Jan 23, 2014·Proceedings of the National Academy of Sciences of the United States of America·Kazuya YamanakaBradley S Moore
Aug 6, 1999·Gene Therapy·G Vassaux
Apr 23, 2008·Current Protocols in Human Genetics·Natalay Kouprina, Vladimir Larionov
Jun 23, 2020·Annual Review of Biochemistry·Weimin ZhangJef D Boeke
Sep 26, 2020·Frontiers in Bioengineering and Biotechnology·Till TisoLars M Blank
Dec 17, 2019·Chemical Reviews·Aleksandra NivinaChaitan Khosla
Aug 4, 2018·ACS Synthetic Biology·Natalay KouprinaVladimir Larionov
Sep 24, 2021·Biotechnology Journal·Yilin LyuHong Lu

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