Journal:

Article Title: Cloning, overexpression, and characterization of a bacterial Ca 2+ -dependent phospholipase D

doi:

Figure Lengend Snippet: Alignment of PLD sequences from Streptomyces. All of the PLDs except that from S. chromofuscus are Ca2+-independent. Residues in red are conserved in all PLDs; residues in blue are conserved among the Ca2+-independent PLDs.

Article Snippet: Cloning of S. chromofuscus (ATCC #23616) pld gene into pET-23a(+) Two oligonucleotide primers were designed based on the published S. chromofuscus PLD DNA sequence ( Yoshioka et al. 1991 ).

Techniques:

Journal:

Article Title: Cloning, overexpression, and characterization of a bacterial Ca 2+ -dependent phospholipase D

doi:

Figure Lengend Snippet: Summary of the purification of PLD fromS. chromofuscus (ATCC strain) cloned from pET-23a(+) and IMPACT systems

Article Snippet: Cloning of S. chromofuscus (ATCC #23616) pld gene into pET-23a(+) Two oligonucleotide primers were designed based on the published S. chromofuscus PLD DNA sequence ( Yoshioka et al. 1991 ).

Techniques: Purification, Clone Assay

Journal:

Article Title: Cloning, overexpression, and characterization of a bacterial Ca 2+ -dependent phospholipase D

doi:

Figure Lengend Snippet: Sequence of S. chromofuscus rPLD with differences from the previously published sequence for S. chromofuscus PLD indicated on the line below. N-PLD would have the first four residues as TTGT instead of ADQA.

Article Snippet: Cloning of S. chromofuscus (ATCC #23616) pld gene into pET-23a(+) Two oligonucleotide primers were designed based on the published S. chromofuscus PLD DNA sequence ( Yoshioka et al. 1991 ).

Techniques: Sequencing

Journal: Methods in enzymology

Article Title: Recombineering: Highly Efficient in vivo Genetic Engineering using Single-strand Oligos

doi: 10.1016/B978-0-12-420067-8.00010-6

Figure Lengend Snippet: TMG

Article Snippet: Primers (see Theory section and Step 1 for design of primers) list-behavior=simple prefix-word= mark-type=none max-label-size=0 Bacto-tryptone Sodium chloride (NaCl) Yeast extract Tris base Magnesium sulfate (MgSO 4 ) Gelatin Bacto Agar Agarose DNA molecular weight markers Ethidium bromide Platinum® Taq DNA Polymerase High Fidelity kit (Invitrogen) (or similar DNA polymerase with proofreading ability) dNTP set Double-distilled sterile chilled H 2 O Recombineering-proficient cells.

Techniques: Concentration Assay