Luciferase genes cloned from the unculturable luminous bacteroid symbiont of the Caribbean flash-light fish,Kryptophanaron alfredi

Margo Haygood, Daniel H. Cohn

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Light organs of anomalopid (flashlight) fish contain luminous bacteroids that have never been cultured and, consequently,have been difficult to study.We have characterized the luciferase (lux)region of DNA extracted from light organs of the Caribbean flashlight fish Kryptophanaron alfredi by hybridization of cloned Vibrio harveyi luxgenes to restriction-endonuclease-digested,light organ DNA. Comparison of the hybridization pattern of light organ DNA with that of DNA of a putative symbiotic isolate provides a method for identifying the authentic luminous symbiont regardless of its luminescence, and was used to reject one such isolate.Light organ DNA was further used to construct a cosmid clone bank and the luciferase genes were isolated.Unlike other bacterial luciferase genes, the genes were not expressed in Escherichia coli.When placed under the control of the E.coli trppromoter,the genes were transcribed but no luciferase was detected,suggesting a posttranscriptional block to expression.

Original languageEnglish (US)
Pages (from-to)203-209
Number of pages7
JournalGene
Volume45
Issue number2
DOIs
StatePublished - 1986
Externally publishedYes

Fingerprint

Luciferases
Fishes
Light
DNA
Genes
Bacterial Luciferases
Escherichia coli
Bacterial Genes
Cosmids
Vibrio
DNA Restriction Enzymes
Luminescence
Clone Cells

Keywords

  • anomalopid
  • bioluminescence
  • clone bank
  • cosmid
  • E.coli
  • gene expression
  • Recombinant DNA
  • transcription
  • Vibrio harveyi

ASJC Scopus subject areas

  • Genetics

Cite this

Luciferase genes cloned from the unculturable luminous bacteroid symbiont of the Caribbean flash-light fish,Kryptophanaron alfredi. / Haygood, Margo; Cohn, Daniel H.

In: Gene, Vol. 45, No. 2, 1986, p. 203-209.

Research output: Contribution to journalArticle

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N2 - Light organs of anomalopid (flashlight) fish contain luminous bacteroids that have never been cultured and, consequently,have been difficult to study.We have characterized the luciferase (lux)region of DNA extracted from light organs of the Caribbean flashlight fish Kryptophanaron alfredi by hybridization of cloned Vibrio harveyi luxgenes to restriction-endonuclease-digested,light organ DNA. Comparison of the hybridization pattern of light organ DNA with that of DNA of a putative symbiotic isolate provides a method for identifying the authentic luminous symbiont regardless of its luminescence, and was used to reject one such isolate.Light organ DNA was further used to construct a cosmid clone bank and the luciferase genes were isolated.Unlike other bacterial luciferase genes, the genes were not expressed in Escherichia coli.When placed under the control of the E.coli trppromoter,the genes were transcribed but no luciferase was detected,suggesting a posttranscriptional block to expression.

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