Bacterial vector for expression of N-terminally 6xHis-tagged proteins with a thrombin site. For other reading frames, use pET-28b (+) or pET-28c (+). Sequence Author: MilliporeSigma …

Feb 12, 2018 · To express recombinant target protein, the pET28a-HMPREF0351_11084 / BL21 (DE3) cells were cultivated in Luria Bertani broth containing kanamycin antibiotic and induced with IPTG. The protein...

Oct 5, 2021 · Experiment 1: Plasmid(pET28a) Isolation from transformed DH5-Alpha cells Aim: To isolate plasmid DNA(pET-28a) from transformed DH5-Alpha cells and visualise the plasmid on 1% agarose gel. Method: 5mL of inoculate was transferred from each of the 4 falcon tubes. where 1,2,3 were the study groups and 4 was the negative control Observations:

Agarose gel electrophoresis of pET-gel and pET-28a(+) plasmids cleaved by double enzymatic hydrolysis. (1) DNA marker; (2) pET-gel digested with both SalI and NdeI; (3) Vector pET28a...

Gel Extraction of the Digested Bands: pET28a/EC2.4.1.5/EC2.4.1.9 > Transformation T4 DNA ligase-mediated ligation of pET28a-2.4.1.5 and pET28a-2.4.1.9

pET28a-Backbone is a commonly used fusion protein type prokaryotic expression vector that contains anti-kanamycin genes and is induced by T7 RNA polymerase provided by host cells. E. coli DH5α and BL21(DE3) are commonly used as the receptor strain.

With the help of BamHI and PflFI restriction enzymes, the sequence of the chimeric protein was put into the pET-28a (+) plasmid. The key benefit of utilising pET expression system is the highly...

Xho1 and Nde and check the size of the digestion products on an agarose gel to confirm that your vector and insert are the expected size in base pairs (bp). As denoted in the vector map, the pET28a vector (minus the segment between the Nde1 and Xho1 sites) is 5,289 bp, and the ABL insert should be 849 bp. As experimental controls, you will

pET 28a Plasmid DNA is a versatile cloning vector that allows for efficient expression and purification of recombinant proteins in E. coli.

We present improved designs and demonstrate that, when incorporated into pET28a, they support increases in protein production. The improved designs are applicable to most of the 103 vectors in the pET series and can be easily implemented.