NDLI: Cloning of human centromeres by transformation-associated recombination in yeast and generation of functional human artificial chromosomes

Content Provider	PubMed Central
Author	Kouprina, N. Ebersole, T. Koriabine, M. Pak, E. Rogozin, I. B. Katoh, M. Oshimura, M. Ogi, K. Peredelchuk, M. Solomon, G. Brown, W. Barrett, J. C. Larionov, V.
Copyright Year	2003
Abstract	Human centromeres remain poorly characterized regions of the human genome despite their importance for the maintenance of chromosomes. In part this is due to the difficulty of cloning of highly repetitive DNA fragments and distinguishing chromosome-specific clones in a genomic library. In this work we report the highly selective isolation of human centromeric DNA using transformation-associated recombination (TAR) cloning. A TAR vector with alphoid DNA monomers as targeting sequences was used to isolate large centromeric regions of human chromosomes 2, 5, 8, 11, 15, 19, 21 and 22 from human cells as well as monochromosomal hybrid cells. The alphoid DNA array was also isolated from the 12 Mb human mini-chromosome ΔYq74 that contained the minimum amount of alphoid DNA required for proper chromosome segregation. Preliminary results of the structural analyses of different centromeres are reported in this paper. The ability of the cloned human centromeric regions to support human artificial chromosome (HAC) formation was assessed by transfection into human HT1080 cells. Centromeric clones from ΔYq74 did not support the formation of HACs, indicating that the requirements for the existence of a functional centromere on an endogenous chromosome and those for forming a de novo centromere may be distinct. A construct with an alphoid DNA array from chromosome 22 with no detectable CENP-B motifs formed mitotically stable HACs in the absence of drug selection without detectable acquisition of host DNAs. In summary, our results demonstrated that TAR cloning is a useful tool for investigating human centromere organization and the structural requirements for formation of HAC vectors that might have a potential for therapeutic applications.
Starting Page	922
File Format	PDF
ISSN	13624962
e-ISSN	13624962
Journal	Nucleic Acids Research
Issue Number	3
Volume Number	31
Language	English
Publisher	Oxford University Press
Publisher Date	2003-02-01
Access Restriction	Open
Rights Holder	Oxford University Press
Subject Keyword	Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Genetics

Sl.	Authority	Responsibilities	Communication Details
1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
5	Website/Portal (Helpdesk)	Queries regarding NDLI and its services	support@ndl.gov.in
6	Contents and Copyright Issues	Queries related to content curation and copyright issues	content@ndl.gov.in
7	National Digital Library of India Club (NDLI Club)	Queries related to NDLI Club formation, support, user awareness program, seminar/symposium, collaboration, social media, promotion, and outreach	clubsupport@ndl.gov.in
8	Digital Preservation Centre (DPC)	Assistance with digitizing and archiving copyright-free printed books	dpc@ndl.gov.in
9	IDR Setup or Support	Queries related to establishment and support of Institutional Digital Repository (IDR) and IDR workshops	idr@ndl.gov.in

Cloning of human centromeres by transformation‐associated recombination in yeast and generation of functional human artificial chromosomes

Specific cloning of human DNA as yeast artificial chromosomes by transformation-associated recombination.

Specific cloning of human DNA as yeast artificial chromosomes by transformation-associated recombination.

Highly selective isolation of human DNAs from rodent–human hybrid cells as circular yeast artificial chromosomes by transformation-associated recombination cloning

Direct isolation of human BRCA2 gene by transformation-associated recombination in yeast.

Highly selective isolation of human DNAs from rodent-human hybrid cells as circular yeast artificial chromosomes by transformation-associated recombination cloning.

Rapid cloning of mouse DNA as yeast artificial chromosomes by transformation-associated recombination (TAR)

Generation of deletion derivatives by targeted transformation of human-derived yeast artificial chromosomes.

Generation of deletion derivatives by targeted transformation of human-derived yeast artificial chromosomes.

Cloning of human centromeres by transformation-associated recombination in yeast and generation of functional human artificial chromosomes

Similar Documents

Cloning of human centromeres by transformation‐associated recombination in yeast and generation of functional human artificial chromosomes

Specific cloning of human DNA as yeast artificial chromosomes by transformation-associated recombination.

Specific cloning of human DNA as yeast artificial chromosomes by transformation-associated recombination.

Highly selective isolation of human DNAs from rodent–human hybrid cells as circular yeast artificial chromosomes by transformation-associated recombination cloning

Direct isolation of human BRCA2 gene by transformation-associated recombination in yeast.

Highly selective isolation of human DNAs from rodent-human hybrid cells as circular yeast artificial chromosomes by transformation-associated recombination cloning.

Rapid cloning of mouse DNA as yeast artificial chromosomes by transformation-associated recombination (TAR)

Generation of deletion derivatives by targeted transformation of human-derived yeast artificial chromosomes.

Generation of deletion derivatives by targeted transformation of human-derived yeast artificial chromosomes.

Cloning of human centromeres by transformation-associated recombination in yeast and generation of functional human artificial chromosomes