Product Data Sheet

Recovery

1. Obtain an LB agar plate with the appropriate antibiotic.

2. Using a sterile pipette tip, touch the bacteria growing within the punctured area of the stab culture. (A sterilized wire loop or sterile toothpick can be used in place of a sterile pipette tip.)

3. Run this tip lightly over a section of the plate, as shown in the figure, to create streak #1.

4. Using another sterile pipette tip, pass through streak #1 and spread the bacteria over a second section of the plate, to create streak #2.

5. Using a third sterile pipette tip, pass through streak #2 and spread the bacteria over the last section of the plate, to create streak #3.

6. Grow overnight in a 37 ^o C incubator (unless a different growth temperature is indicated on the plasmid datasheet).

7. In the morning, single colonies should be visible. If the bacterial growth is too dense, re-streak onto a new agar plate to obtain single colonies.

LuxCDABE sequence in pBBR1MCS4-LuxCDABE

Photorhabdus luminescens fatty acid reductase (luxC), acyltransferase (luxD), luciferase (luxA), luciferase (luxB), and luxE (luxE) genes, complete cds

GenBank: M90093.1

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LOCUS       XENLABCDEA              6960 bp    DNA     linear   BCT 11-APR-2001

DEFINITION  Photorhabdus luminescens fatty acid reductase (luxC),

            acyltransferase (luxD), luciferase (luxA), luciferase (luxB), and

            luxE (luxE) genes, complete cds.

ACCESSION   M90093

VERSION     M90093.1  GI:155405

KEYWORDS    .

SOURCE      Photorhabdus luminescens subsp. luminescens

  ORGANISM  Photorhabdus luminescens subsp. luminescens

            Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacteriales;

            Enterobacteriaceae; Photorhabdus.

REFERENCE   1  (bases 1 to 6960)

  AUTHORS   Meighen,E.A. and Szittner,R.B.

  TITLE     Multiple repetitive elements and organization of the lux operons of

            luminescent terrestrial bacteria

  JOURNAL   J. Bacteriol. 174 (16), 5371-5381 (1992)

   PUBMED   1644764

FEATURES             Location/Qualifiers

     source          1..6960

                     /organism="Photorhabdus luminescens subsp. luminescens"

                     /mol_type="genomic DNA"

                     /strain="Hb"

                     /sub_species="luminescens"

                     /db_xref="taxon:171439"

                     /tissue_lib="ATCC 29999"

     gene            459..1901

                     /gene="luxC"

     CDS             459..1901

                     /gene="luxC"

                     /codon_start=1

                     /transl_table=11

                     /product="fatty acid reductase"

                     /protein_id="AAA27617.1"

                     /db_xref="GI:155406"

                     NIIDCNGNNELRLHNIVNFLYTVGQRWKNEEYSRRRTYIRDLKKYMGYSEEMAKLEAN

                     WISMILCSKGGLYDVVENELGSRHIMDEWLPQDESYVRAFPKGKSVHLLAGNVPLSGI

                     MSILRAILTKNQCIIKTSSTDPFTANALALSFIDVDPNHPITRSLSVIYWPHQGDTSL

                     AKEIMRHADVIVAWGGPDAINWAVEHAPSYADVIKFGSKKSLCIIDNPVDLTSAATGA

                     AHDVCFYDQRACFSAQNIYYMGNHYEEFKLALIEKLNLYAHILPNAKKDFDEKAAYSL

                     VQKESLFAGLKVEVDIHQRWMIIESNAGVEFNQPLGRCVYLHHVDNIEQILPYVQKNK

                     TQTISIFPWESSFKYRDALALKGAERIVEAGMNNIFRVGGSHDGMRPLQRLVTYISHE

                     RPSNYTAKDVAVEIEQTRFLEEDKFLVFVP"

     gene            1913..2836

                     /gene="luxD"

     CDS             1913..2836

                     /gene="luxD"

                     /codon_start=1

                     /transl_table=11

                     /product="acyltransferase"

                     /protein_id="AAA27618.1"

                     /db_xref="GI:155407"

                     /translation="MENESKYKTIDHVICVEGNKKIHVWETLPEENSPKRKNAIIIAS

                     GFARRMDHFAGLAEYLSRNGFHVIRYDSLHHVGLSSGTIDEFTMSIGKQSLLAVVDWL

                     TTRKINNFGMLASSLSARIAYASLSEINASFLITAVGVVNLRYSLERALGFDYLSLPI

                     NELPDNLDFEGHKLGAEVFARDCLDFGWEDLASTINNMMYLDIPFIAFTANNDNWVKQ

                     DEVITLLSNIRSNRCKIYSLLGSSHDLSENLVVLRNFYQSVTKAAIAMDNDHLDIDVD

                     ITEPSFEHLTIATVNERRMRIEIENQAISLS"