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Peptide-coding sequence

Overexpression of apoaequorin (Inouye et al., 1989, 1991). To produce a large quantity of apoaequorin, an apoaequorin expression plasmid piP-HE containing the signal peptide coding sequence of the outer membrane protein A (ompA) of E. coli (Fig. 4.1.12) was constructed and expressed in E. coli. The expressed apoaequorin was secreted into the periplasmic space of bacterial cells and culture medium. The cleaving of ompA took place during secretion thus the... [Pg.116]

Figure 1 Strategy for cloning a peptide-coding sequence (CDS) as tandem repeats in the vector pET31b. The resulting fusion protein, comprising the ketosteroid isomerase (KSI), peptide repeats, and His-tag, is targeted to inclusion bodies. The fusion protein can be recovered and cleaved, in this case, with cyanogen bromide (CNBr) which acts at the methionine (M) residues allowing further separation of pure peptide from the other fusion components. The cleavage by CNBr results in a C-terminal homoserine lactone (hsl) on each peptide monomer. Figure 1 Strategy for cloning a peptide-coding sequence (CDS) as tandem repeats in the vector pET31b. The resulting fusion protein, comprising the ketosteroid isomerase (KSI), peptide repeats, and His-tag, is targeted to inclusion bodies. The fusion protein can be recovered and cleaved, in this case, with cyanogen bromide (CNBr) which acts at the methionine (M) residues allowing further separation of pure peptide from the other fusion components. The cleavage by CNBr results in a C-terminal homoserine lactone (hsl) on each peptide monomer.
The host cell used in this experiment is Epicurian coli XL 1-Blue. This strain contains a deletion on the chromosome of the first 450 base pairs of its lacZ coding sequence (the LacZa peptide). The pUC19 plasmid will be able to restore /3-galac-tosidase activity in the host strain, since this plasmid contains an uninterrupted LacZa peptide coding sequence (the strain will be complemented for /3-galactosidase activity). The LacZa peptide expressed from the pUC19 plasmid will be able to... [Pg.349]

These last factors also up-regulate hBD-2 secretion by the intestinal epithelial cell lineage Caco-2 stimulated with probiotic bacteria [176]. Although not yet demonstrated experimentally, mammalian cathelicidins may respond to inflammatory stimuli, since the 5 flanking sequences upstream of the peptide coding sequence have several potential consensus sequences for transcription factors involved in inflammatory response, sueh as NF-kB, NF-IL-6, acute-phase response factor and IFN-y response elements [147,170,177]. [Pg.639]

Steenbergh PH, Hoppener JW, Zandberg J, et al. Calcitonin gene related peptide coding sequence is conserved in the human genome and is expressed in medullary thyroid carcinoma. J Clin Endocrinol Metab. 1984 59 358-360. [Pg.333]

We wanted to know if all genes in one isozyme family responded similarly to GA. In the case of the low-pl genes this answer was relatively easy to approach. Primer extension experiments were performed. In these experiments [26] a DNA fragment that could hybridize to the signal peptide coding sequence of the low-pl cDNA was labeled with at one end, allowed to hybridize to RNA isolated from... [Pg.135]

The results from similar primer extension experiments using a labeled primer specific for the high-pl cDNA signal peptide coding sequence were more difficult to explain. In these experiments, all primer extension products identified had the same sequence for a distance 50 nucleotides in the 5 direction from the ATG translation initiation codon (it was not possible to obtain sequence data further... [Pg.135]

The coding sequences of a- and P-globins of human hemoglobin have been fused to the sequence of the chloroplastic transit peptide of the small subunit of Rubisco. These proteins were then coexpressed in transgenic tobacco plants, resulting in the production of a functional form of tetra-meric hemoglobin. The results demonstrate that a complex multimeric protein such as recombinant human hemoglobin can be obtained from tobacco in a functional form. [Pg.43]

The last stage of peptide chain synthesis is termination. The genetic code specifies three stop codons, indicating the termination of a coding sequence. When the ribosome encounters one of these stop codons on the mRNA, certain release factors... [Pg.21]

Over several decades, multiple vector systems for recombinant gene expression in E. coli have been developed. Modem vectors suitable for recombinant protein production vary in the used promoter system in the presence or absence of coding sequences for affinity tags upstream or downstream of the multiple cloning site (MCS) and of sequences coding for leader peptides for the protein export. Moreover, different origins of replication (ori), antibiotic selection marker genes and MCS are used. [Pg.136]


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Coding sequence

Peptide sequences

Peptide sequencing

Peptidic sequences

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