INTEGRATIVE SELECTION OF HUMAN CHROMOSOME-SPECIFIC YEAST ARTIFICIAL CHROMOSOMES

被引：9

作者：

PAVAN, WJ ^{[1
]}

REEVES, RH ^{[1
]}

机构：

[1] JOHNS HOPKINS UNIV,SCH MED,DEPT PHYSIOL,DEV GENET LAB,725 N WOLFE ST,BALTIMORE,MD 21205

来源：

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA | 1991年 / 88卷 / 17期

关键词：

YEAST ARTIFICIAL CHROMOSOME LIBRARY; HOMOLOGOUS RECOMBINATION; GENETIC VECTOR;

D O I：

10.1073/pnas.88.17.7788

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Human specific "integrative selection vectors" (ISVs) were designed to optimize integration of a yeast-selectable marker specifically into yeast artificial chromosomes (YACs) derived from human but not mouse DNA. ISVs were transformed into a YAC genomic library constructed from DNA of a human-mouse somatic cell hybrid containing chromosome 21 (HSA21) as the only human chromosome. One percent of the yeast in the original library contained HSA21-derived YACs; between 45% and 54% of the yeast recovered after transformation with ISV vectors contained human YACs. Integrative selection provides a rapid means of obtaining a highly enriched population of human chromosome-specific YACs by eliminating the labor-intensive steps of isolating and screening primary transformants. The procedure is biased toward the selection of YACs that contain a large number of targets for homologous recombination: thus, libraries constructed by this procedure will be composed primarily of the largest YACs in the population.

引用

页码：7788 / 7791

页数：4

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