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- PBluescript abstract "In genetics, pBluescript (pBS) or pBluescript II is a commercially available phagemid containing several useful sequences for use in cloning with bacteriophage. The sequences include a multiple cloning site sequence (MCS), antibiotic resistance sequence to ampicillin and an E. coli and f1 helper phage origin of replication. The multiple cloning site sequence is located within a LacZ controlled gene designed to provide a blue coloration when expressed in bacteria. This is usually achieved via X-gal found in agarose growth media used to culture bacteria with pBS. If the gene is disrupted by successful insertion of a DNA sequence, the bacteria exhibit a white coloration in Blue white screening, distinguishing successful recombination from those phagemids which were not altered. These recombinant plasmids can then be used in a variety of molecular techniques. Determining expression and developing genomic libraries are some applications for which the pBS recombinants can be used.The pBluescript II phagemids (plasmids with a phage origin) are cloningvectors designed to simplify commonly used cloning and sequencingprocedures, including the construction of nested deletions for DNAsequencing, generation of RNA transcripts in vitro and site-specificmutagenesis and gene mapping. The pBluescript II phagemids have anextensive multiple cloning site with 21 unique restriction enzyme recognition sites.Flanking the multiple cloning site are T7 and T3 RNA polymerase promoters that canbe used to synthesize RNA in vitro.1, 2The choice of promoter used to initiate transcription determines which strand of the insert cloned into themultiple cloning site will be transcribed. Circular maps and lists of features for the pBluescript II phagemids areshown in figures 1 and 2. The multiple cloning site and T7 and T3 RNA polymerasepromoter sequences are present in the N-terminal portion of a lacZ genefragment. A total of 131 amino acids of β-galactosidase coding sequence ispresent in the pBluescript II phagemid, but the coding sequence isinterrupted by the large multiple cloning site. (There are 36 amino acids from theinitiator Met sequence to the EcoR I site.) pBluescript II phagemids havingno inserts in the multiple cloning site will produce blue colonies in the appropriatestrains of bacteria (i.e., strains containing lacZΔM15 on an F´ episome, suchas XL1-Blue MRF´, among others). pBluescript II phagemids that haveinserts will produce white colonies using the same strain, because the insertsdisrupt the coding region of the lacZ gene fragment.pBluescript II (+) and (–) are available with two multiple cloning site orientationsdesignated as either KS or SK using the following convention: (1) in the KSorientation, the Kpn I restriction site is nearest the lacZ promoter and theSac I restriction site is farthest from the lacZ promoter; and (2) in the SKorientation, the Sac I site is the closest restriction site to the lacZ promoterand the Kpn I site is the farthest. Flanking the T3 and T7 promoters are BssH II sites. This rare six-base cutterwill allow the insert plus the T phage RNA promoters to be excised andused for gene mapping.pBluescript II phagemids can be rescued as single-stranded (ss) DNA.pBluescript II phagemids contain a 454-bp filamentous f1 phage intergenicregion (M13 related), which includes the 307-bp origin of replication. The(+) and (–) orientations of the f1 intergenic region allow the rescue of senseor antisense ssDNA by a helper phage. This ssDNA can be used fordideoxynucleotide sequencing (Sanger method) or site-specific mutagenesis.".
- PBluescript wikiPageID "3755776".
- PBluescript wikiPageRevisionID "572466536".
- PBluescript hasPhotoCollection PBluescript.
- PBluescript subject Category:Genetics.
- PBluescript subject Category:Plasmids.
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- PBluescript type NaturalObject100019128.
- PBluescript type Object100002684.
- PBluescript type PhysicalEntity100001930.
- PBluescript type Plasmid109395266.
- PBluescript type Plasmids.
- PBluescript type Whole100003553.
- PBluescript comment "In genetics, pBluescript (pBS) or pBluescript II is a commercially available phagemid containing several useful sequences for use in cloning with bacteriophage. The sequences include a multiple cloning site sequence (MCS), antibiotic resistance sequence to ampicillin and an E. coli and f1 helper phage origin of replication. The multiple cloning site sequence is located within a LacZ controlled gene designed to provide a blue coloration when expressed in bacteria.".
- PBluescript label "PBluescript".
- PBluescript label "PBluescript".
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- PBluescript sameAs Q7118460.
- PBluescript sameAs Q7118460.
- PBluescript sameAs PBluescript.
- PBluescript wasDerivedFrom PBluescript?oldid=572466536.
- PBluescript isPrimaryTopicOf PBluescript.