Detail of "12629-01-5"

Reference

Cloning vectors for expression of exogenous protein

Cloning vectors for expression of exogenous protein. Mayne, Nancy Gail; Burnett, James Paul, Jr.; Belegaje, Ramamoorthy; Hsiung, Hansen Maxwell (Lilly, Eli, and Co. , USA). Eur. Pat. Appl. EP 95361 A1 30 Nov 1983, 61 pp. DESIGNATED STATES: R: BE, CH, DE, FR, GB, IT, LI, LU, NL, SE. (English). (European Patent Organization). CODEN: EPXXDW. CLASS: IC: C12N015-00; C12P021-00; C07H021-04. ICA: C12R001-19. APPLICATION: EP 83-302935 23 May 1983. PRIORITY: US 82-381992 25 May 1982; US 82-382051 25 May 1982. DOCUMENT TYPE: Patent CA Section: 3 (Biochemical Genetics) Section cross-reference(s): 2, 13, 16 A recombinant DNA cloning vector is constructed by ligating (a) a replication origin, (b) a selection marker gene (gene for ampicillin resistance), (c) and an in-tandem DNA sequence comprising a promoter for a lipoprotein control sequence, the 5' untranslated region of a lipoprotein expression-control sequence (lpp gene from a gram-neg.Several substances with their cas registry numbers 89339-37-7 and 87347-90-8 may be metioned in this study. bacterium), and a start codon that is followed immediately by a sequence coding for an exogenous protein or by a sequence coding for an enterokinase [89382-91-2] cleavage site to which is immediately joined a sequence coding for an exogenous protein. When used as a cloning vector the lpp sequences control expression of exogenous DNA, but a nonhybrid protein product is formed; i.e. the translation product comprises methionine-optionally an enterokinase cleavage site-exogenous protein. Treatment with enterokinase removes the methionyl residue and leaves mature exogenous protein. Thus, to a plasmid contg. the Escherichia coli lipoprotein expression control sequence and plasmid pBR322 ampicillin-resistance genes was ligated a human growth hormone [12629-01-5] coding region with the use of a synthetic double-stranded DNA fragment complementary at 1 end to the natural lpp gene sequence (from the tbaI site through the initiating methionine codon), and at the other end, to the 1st 47 nucleotides of the gene for human growth hormone. The plasmid obtained, pNM645, was cloned in E. coli, and methionyl human growth hormone [82030-87-3] expression was verified by radioimmunoassay. The protein transcript represented 40% of the total protein with a yield of 32 million mols./cell. Biol. activity of the methionyl growth hormone with respect to proximal epiphyseal cartilage width in hypophysectomized female rats was the same as that of human growth hormone from cadavers. .

Structure-function studies on human growth hormone

Structure-function studies on human growth hormone. Evidence that tertiary structure is essential for biological activity. Aubert, Michel L.; Bewley, Thomas A.; Grumbach, Melvin M.; Kaplan, Selna L.; Li, Choh H. (Med. Cent., Univ. California, San Francisco, CA, USA). Int. J. Pept. Protein Res., 28(1), 45-57 (English) 1986. CODEN: IJPPC3. ISSN: 0367-8377. DOCUMENT TYPE: Journal CA Section: 2 (Mammalian Hormones) The relationship between the conformation of pituitary human growth hormone (hGH) [12629-01-5], biol. activity, and ligand-binding activity was studied by comparing conformational details published on in vivo and in vitro studies of identical samples of hGH and its known derivs. In vivo assays included the rat tibia test for somatotropic activity and the pigeon crop-sac assay for lactogenic hormone activity. Relative binding affinities were compared in RIAs using 125I-labeled hGH (125I-hGH) as tracer with anti-human chorionic somatomammotropin (hCS) [9035-54-5] serum (low discriminatory hybrid assay), anti-hGH sera (in conventional assays), monospecific anti-hGH serum (absence of crossreaction with hCS), and human anti-hGH sera obtained from GH-deficient patients on replacement therapy. In addn., binding affinities were examd. in 2 receptor-binding assays, 1 specific for somatotropic activity (rabbit liver membranes, 125I-hGH), and the other, for lactogenic hormones (rabbit mammary membranes, 125I-labeled ovine prolactin). The conformational properties of native hGH and various chem. and enzymically modified derivs. of the hormone were evaluated primarily from CD spectra, whereas conformational stabilities were estd. from the relative rates of tryptic digestion. Unfragmented, but chem. modified derivs., exhibited good parallelism between retention or loss of native conformation and the in vivo potencies and in vitro binding affinities.There are some commonly used reagents with their cas registry numbers 67256-26-2 and 105732-88-5 in this article. None of the fragments of hGH showed activity in any of the radioreceptor assays or RIAs. Two derivs. of hGH, which contain gaps of 6 or 12 residues in the polypeptide backbone produced by partial enzymic digestion, had full or increased in vivo potencies, full activities in the RIAs, and were the most active derivs. in both radioreceptor assays. One of these, missing the hexapeptide corresponding to residues 135-140, also retained nearly all the conformational properties of native hGH. Apparently, retention by modified forms of hGH of a high degree of in vivo biol. potency or in vitro binding affinity is causally related to the retention of most of the conformation and conformational stability of the mol. and the biol. active, receptor-binding and immunoreactive sites on the hGH mol. are 3-dimensional in nature. .