- Function: The Hind III sites in pBR322 are specific sequences where the Hind III enzyme can cleave the DNA. This cleavage is essential for inserting foreign DNA into the plasmid, making it a vital tool in genetic engineering. The ability to cut DNA at specific sites allows for the manip...
The cleavage depends on the substrate used: it varies from the near complete digestion of two TspGWI sites in pUC19 DNA (Figure 9B, lane 2) to the trace cleavage of a subset of refractive sites in pBR322 and pACYC184 plasmids [10]. The refractory site phenomenon was preliminary evaluated ...
In order to clone the 462 bp Alu I fragment containing the lpp promoter region in pBR322, the DNA fragment lying between the Eco RI and Hind III cleavage sites of pBR322 (containing the promoter of the tetracycline resistance gene) was first deleted, as shown schematically at 106 in FIG....
The scheme of theygjGgene andaer-ygjGintergenic region. A. The fragment (from -311 to +1480) containing ygjG gene (b3073) with aer-ygjG intergenic region, and sites for restriction enzymes used in cloning procedure are shown. The orientation of genes and location of σ54promoter are repr...
10. The vector of claim 9 in which the cloning site is SnaB I. 11. The vector of claim 10 in which the Sfi I and Not I sites flanking the SUP4 gene are deleted. 12. The vector of claim 10 in which an EcoR I linker is inserted into the SnaB I site. ...
Plasmid pLA2901 harbors genes coding for resistance to kanamycin and tetracycline. It contains 5 unique restriction sites: the EcoRl site of pRK290, the HindIII site of pBR322, and the BglII, BstEII, and XhoI sites of Tn5. DNA insertions at the unique BglII site can be detected by inac...
the O gene (O/2). The pKC30 plasmid is a derivative of plasmid pBR322 which contains a HindIII-BamHI restriction fragment derived from phage λ inserted between the HindIII and BamHI restriction sites within the tetracycline gene of pBR322. The λ insert contains the promoter signal, PL,...
yeast and viruses. The origin of replication from the plasmid pBR322 is suitable for most Gram-negative bacteria, the 2m plasmid origin is suitable for yeast, and various viral origins (SV40, polyoma, adenovirus, VSV or BPV) are useful for cloning vectors in mammalian cells. Generally, the...
appropriate cleavage sites are coded for adjacent the polypeptide-additional protein codon junction. Thus, in FIG. 1 as an example, the expression product is a precursor protein comprising both somatostatin and the greatest part of the β-galactosidase polypeptide. Here ATG is not required to code...
Human pancreatic islet cell glutamic acid decarboxylase (GAD), an autoantigen involved in the development of insulin-dependent diabetes mellitus (IDDM), has been cloned, sequenced and expressed by rec