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| Vectors | | |  | pET-44
The pET-44 vectors are designed for cloning and high-level expression of polypeptide sequences fused with the 495 aa NusA (Nus•Tag™) protein. The solubility of many target proteins in the E. coli cytoplasm is increased when they are expressed NusA fusions. The pET-44a-c(+) vectors, which are derived from the corresponding pET-43.1 series, encode an additional N-terminal His•Tag® sequence for enhanced yields when purifying a target protein with His•Mag™ Agarose Beads from lysates generated with the BugBuster® HT Reagent and PopCulture® Reagent. N-terminal S•Tag™ and optional C-terminal HSV•Tag® and His•Tag fusion sequences are also available for versatile detection and purification of target proteins.
pET-43.1
The pET-43.1 series of vectors are designed for cloning and high-level expression of peptide sequences fused with the 491 aa Nus•Tag™ protein. Note that the sequence is numbered by the pBR322 convention, so the T7 expression region is reversed on the vector map (TB288). The f1 origin is oriented so that infection with helper phage will produce virions containing single stranded DNA that corresponds to the coding strand. Therefore, single stranded sequencing should be performed using the ColiDOWN primer. Vector encoded sequence can be completely removed when cloning into the PshA I or Sma I sites by cleaving the Nus•Tag fusion protein with Enterokinase or Thrombin, respectively.
pET-42
The pET-42 series is designed for cloning and high-level expression of peptide sequences fused with the 220 aa GST•Tag™ protein. Note that the sequence is numbered by the pBR322 convention, so the T7 expression region is reversed on the vector map (TB240). The f1 origin is oriented so that infection with helper phage will produce virions containing single stranded DNA that corresponds to the coding strand. Therefore, single stranded sequencing should be performed using the T7 terminator primer. Vector encoded sequence can be completely removed when cloning into the PshAI site (as shown below) and then cleaving the GST fusion protein with Factor Xa.
pET-41
The pET-41 series is designed for cloning and high-level expression of peptide sequences fused with the 220 aa GST•Tag™ protein. Note that the sequence is numbered by the pBR322 convention, so the T7 expression region is reversed on the vector map (TB239). The f1 origin is oriented so that infection with helper phage will produce virions containing single stranded DNA that corresponds to the coding strand. Therefore, single stranded sequencing should be performed using the T7 terminator primer.Vector encoded sequence can be completely removed when cloning into the PshAI site (as shown below) and then cleaving the GST fusion protein with Enterokinase.
pET-32
The pET-32a vector is designed for cloning and high-level expression of peptide sequences fused with the 109aa Trx•Tag™ thioredoxin protein (1). Cloning sites are available for producing fusion proteins also containing cleavable His•Tag® and S•Tag™ sequences for detection and purification. Note that the sequence is numbered by the pBR322 convention, so the T7 expression region is reversed on the vector map (TB122 ). The f1 origin is oriented so that infection with helper phage will produce virions containing single-stranded DNA that corresponds to the coding strand. Therefore, single-stranded sequencing should be performed using the T7 terminator primer. |
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