A cosmid with a HyR marker for fungal library construction and screening

Research output: Contribution to journalArticle

123 Citations (Scopus)

Abstract

The construction of a double-cos-site cosmid vector, pMOcosX, for use in making filamentous fungal genomic DNA libraries, is described. The vector has features that allow for selection of clones introduced into fungi by transformation and for efficient chromosome walking experiments. These features include (i) two cos sites allowing for easy construction of libraries without requiring size selection of insert DNA; (ii) an XhoI site for insertion of Sau3AI or MboI partially digested genomic DNA inserts that allows usage of a half-site fill-in method which minimizes the possibility of producing clones containing chimeric inserts; (iii) a bacterial hygromycin phosphotransferase-encoding gene fused to a modified cpc-1 promoter of Neurospora crassa for direct selection of cosmid clones upon introduction into fungal cells; and (iv T7 and T3 bacteriophage promoters and EcoRI, NotI and BamHI restriction sites flanking the cloning site that allow for synthesis of, or isolation of, end-specific probes for chromosome walking. The combination of features in this vector allows for the easy construction and use of high-quality fungal DNA libraries from small amounts of genomic DNA.

Original languageEnglish (US)
Pages (from-to)159-162
Number of pages4
JournalGene
Volume150
Issue number1
DOIs
StatePublished - Dec 2 1994
Externally publishedYes

Fingerprint

Cosmids
Chromosome Walking
Fungal DNA
Libraries
hygromycin-B kinase
Clone Cells
Gene Library
DNA
Bacteriophage T3
Bacteriophage T7
Neurospora crassa
Genomic Library
Organism Cloning
Fungi
Genes

Keywords

  • double cos site
  • hygromycin resistance
  • Magnaporthe grisea
  • Neurospora crassa
  • pMOcosX
  • Recombinant DNA
  • T3 phage promoters
  • T7 phage promoters

ASJC Scopus subject areas

  • Genetics

Cite this

A cosmid with a HyR marker for fungal library construction and screening. / Orbach, Marc Joel.

In: Gene, Vol. 150, No. 1, 02.12.1994, p. 159-162.

Research output: Contribution to journalArticle

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