Construction and characterization of a yeast artificial chromosome library containing seven haploid human genome equivalents

Hans M. Albertsen, Hadi Abderrahim, Howard M. Cann, Jean Dausset, Denis L E Paslier, Daniel Cohen

Research output: Contribution to journalArticle

490 Citations (Scopus)

Abstract

Prior to constructing a library of yeast artificial chromosomes (YACs) containing very large human DNA fragments, we performed a series of preliminary experiments aimed at developing a suitable protocol. We found an inverse relationship between YAC insert size and transformation efficiency. Evidence of occasional rearrangement within YAC inserts was found resulting in clonally stable internal deletions or clonally unstable size variations. A protocol was developed for preparative electrophoretic enrichment of high molecular mass human DNA fragments from partial restriction digests and ligation with the YAC vector in agarose. A YAC library has been constructed from large fragments of DNA from an Epstein-Barr virus-transformed human lymphoblastoid cell line. The library presently contains 50,000 clones, 95% of which are greater than 250 kilobase pairs in size. The mean YAC size of the library, calculated from 132 randomly isolated clones, is 430 kilobase pairs. The library thus contains the equivalent of approximately seven haploid human genomes.

Original languageEnglish
Pages (from-to)4256-4260
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume87
Issue number11
Publication statusPublished - 1 Dec 1990
Externally publishedYes

Fingerprint

Yeast Artificial Chromosomes
Haploidy
Human Genome
Libraries
DNA
Clone Cells
Human Herpesvirus 4
Sepharose
Ligation
Cell Line

Keywords

  • Physical mapping
  • Preparative pulsed-field electrophoresis

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Construction and characterization of a yeast artificial chromosome library containing seven haploid human genome equivalents. / Albertsen, Hans M.; Abderrahim, Hadi; Cann, Howard M.; Dausset, Jean; Paslier, Denis L E; Cohen, Daniel.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 87, No. 11, 01.12.1990, p. 4256-4260.

Research output: Contribution to journalArticle

Albertsen, Hans M. ; Abderrahim, Hadi ; Cann, Howard M. ; Dausset, Jean ; Paslier, Denis L E ; Cohen, Daniel. / Construction and characterization of a yeast artificial chromosome library containing seven haploid human genome equivalents. In: Proceedings of the National Academy of Sciences of the United States of America. 1990 ; Vol. 87, No. 11. pp. 4256-4260.
@article{769cc8d4ea0b4e378c3e9334fe887c28,
title = "Construction and characterization of a yeast artificial chromosome library containing seven haploid human genome equivalents",
abstract = "Prior to constructing a library of yeast artificial chromosomes (YACs) containing very large human DNA fragments, we performed a series of preliminary experiments aimed at developing a suitable protocol. We found an inverse relationship between YAC insert size and transformation efficiency. Evidence of occasional rearrangement within YAC inserts was found resulting in clonally stable internal deletions or clonally unstable size variations. A protocol was developed for preparative electrophoretic enrichment of high molecular mass human DNA fragments from partial restriction digests and ligation with the YAC vector in agarose. A YAC library has been constructed from large fragments of DNA from an Epstein-Barr virus-transformed human lymphoblastoid cell line. The library presently contains 50,000 clones, 95{\%} of which are greater than 250 kilobase pairs in size. The mean YAC size of the library, calculated from 132 randomly isolated clones, is 430 kilobase pairs. The library thus contains the equivalent of approximately seven haploid human genomes.",
keywords = "Physical mapping, Preparative pulsed-field electrophoresis",
author = "Albertsen, {Hans M.} and Hadi Abderrahim and Cann, {Howard M.} and Jean Dausset and Paslier, {Denis L E} and Daniel Cohen",
year = "1990",
month = "12",
day = "1",
language = "English",
volume = "87",
pages = "4256--4260",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "11",

}

TY - JOUR

T1 - Construction and characterization of a yeast artificial chromosome library containing seven haploid human genome equivalents

AU - Albertsen, Hans M.

AU - Abderrahim, Hadi

AU - Cann, Howard M.

AU - Dausset, Jean

AU - Paslier, Denis L E

AU - Cohen, Daniel

PY - 1990/12/1

Y1 - 1990/12/1

N2 - Prior to constructing a library of yeast artificial chromosomes (YACs) containing very large human DNA fragments, we performed a series of preliminary experiments aimed at developing a suitable protocol. We found an inverse relationship between YAC insert size and transformation efficiency. Evidence of occasional rearrangement within YAC inserts was found resulting in clonally stable internal deletions or clonally unstable size variations. A protocol was developed for preparative electrophoretic enrichment of high molecular mass human DNA fragments from partial restriction digests and ligation with the YAC vector in agarose. A YAC library has been constructed from large fragments of DNA from an Epstein-Barr virus-transformed human lymphoblastoid cell line. The library presently contains 50,000 clones, 95% of which are greater than 250 kilobase pairs in size. The mean YAC size of the library, calculated from 132 randomly isolated clones, is 430 kilobase pairs. The library thus contains the equivalent of approximately seven haploid human genomes.

AB - Prior to constructing a library of yeast artificial chromosomes (YACs) containing very large human DNA fragments, we performed a series of preliminary experiments aimed at developing a suitable protocol. We found an inverse relationship between YAC insert size and transformation efficiency. Evidence of occasional rearrangement within YAC inserts was found resulting in clonally stable internal deletions or clonally unstable size variations. A protocol was developed for preparative electrophoretic enrichment of high molecular mass human DNA fragments from partial restriction digests and ligation with the YAC vector in agarose. A YAC library has been constructed from large fragments of DNA from an Epstein-Barr virus-transformed human lymphoblastoid cell line. The library presently contains 50,000 clones, 95% of which are greater than 250 kilobase pairs in size. The mean YAC size of the library, calculated from 132 randomly isolated clones, is 430 kilobase pairs. The library thus contains the equivalent of approximately seven haploid human genomes.

KW - Physical mapping

KW - Preparative pulsed-field electrophoresis

UR - http://www.scopus.com/inward/record.url?scp=0025368589&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025368589&partnerID=8YFLogxK

M3 - Article

C2 - 2190217

AN - SCOPUS:0025368589

VL - 87

SP - 4256

EP - 4260

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 11

ER -