• Proceedings of the Korean Society of Toxicology Conference
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• 1997.10a
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• pp.38-42
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• 1997

Human growth hormone is known as one of the peptide hormones which is consisted of 191 amino acids derived from the pituitary gland in humans. The objectives of this study were to supply inexpensive recombinant methionyl human growth hormones (rHGH) synthesized by the DNA technology in a yeast cell line and followed by the establishement of protein purification techniques. The next steps of the research were to study its physic-chemical properties and biological properties, and to evaluate various preclinical aspcts including pharmacokinetics sutdy, general pharmacology study, general toxicity test, and specific toxicity tests. Clinical phase I, II, III studies were also done against growth hormone dficient children to reveal that growth promoting effects were similar compared with the natural HGH extracted from pituitary glands and commercially available rHGHs. The results could be summarized that (I) this yeast dervied rHGH have had excellent physico-chemical and biological properties in comparison with a natural HGH and other synthesized rHGHs, (2) we could not see any toxic side effects when very high doses were administered to the experimental animals, and (3) this growth hormone showed effectiveness in the growth stimulating to growth hormone deficient patients.

• Fisheries and Aquatic Sciences
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• v.6 no.3
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• pp.116-124
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• 2003

Isolation and cloning of seven-band grouper (Epinephelus septemfasciatus) growth hormone cDNA from pituitary gland revealed an open reading frame of 612 bp coding for a pre-growth hormone of 204 amino acids with a 17 amino acid putative signal peptide. Deduced amino acid sequence showed that there was one possible N-glycosylation site at $Asn^{l84}$ and four cysteine residues $(Cys^{52},\;Cys^{160},\;Cys^{177},\;Cys^{185})$ on t e same positions as in some other species where they were involved in the stabilization of the tertiary structure. The seven-band grouper growth hormone (sbgGH) presented a $99.5\%$ amino acid sequence identity with the growth hormone of Epinephelus coioides and contained the conserved hormone domain region. Comparison of growth hormone sequences from evolutionarily diverse species revealed 25 amino acid residues conserved in jawless fishes to modern mammals. It also revealed an evolutionary trend to retain the same polypeptide sequence even in the distantly related animals while allowing alterations to occur in polypeptides of the closely related species. In order to create a recombinant system to produce high levels of the growth hormone, it was expressed in Escherichia coli (BL21) cells. The gel analysis revealed theoretically expected molecular weights for both mature and pre-sbgGHs.

• The Journal of Korean Society of Virology
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• v.28 no.2
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• pp.129-138
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• 1998

Bovine growth hormone (bGH) gene was expressed in an insect Spodoptera frugiperda cell line using a Baculovirus, Hyphantria cunea nuclear polyhedrosis virus (HcNPV). The bGH gene in pbGH plasmid was sequenced and amplified by PCR technique with two primers containing NcoI sites. The bGH gene consisted of 654 bp (217 amino acid residues), the 5'-untranslated region of the cloned bGH cDNA contains 56 bp, and the 3'-untranslated region contains 145 bp and two pallindromic regions. The amplified bGH gene DNA fragment (654 bp) was inserted into the NcoI site of the pHcEVII vector, which was named pHcbGH. The pHcbGH transfer vector DNA and the wild type HcNPV DNA were cotransfected into S. frugiperda cells to construct a recombinant virus. Eight recombinant viruses were selected and named HcbGH. One clone, HcbGH-4-1 showed largest plaque size, therefore the recombinant virus was further studied. The multiplication pattern of the recombinant HcbGH-4-1 was similar to that of the wild type HcNPV. The bGH gene DNA in the HcbGH-4-1 recombinant was confirmed by Southern blot hybridization. The amount of the bGH (217 amino acid residues, 21 kDa) produced in S. frugiperda cells infected with the HcbGH-4-1 recombinant was approximately 5.5 ng per ml ($10^6$ cells) by radioimmunoassay.

• 한국생물공학회:학술대회논문집
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• 2003.04a
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• pp.554-556
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• 2003

This research was focused on expression, purification and characterization of ubiquitin-specific protease 1 (UBP1) expressed in recombinant Escherichia coli. Various systems were constructed by fusing polycationic fusion tails or fusion partners to the C- or N-terminus of the product protein. In particular, UBP1 containing 6 histidine residues at the N-terminal end showed best results in terms of expression level and purification efficiency. The N-terminal $6{\times}His-tagged$ UBP1 was overproduced in recombinant E. coli using high cell density cultivation technology and purified using immobilized metal affinity chromatography. The molecular weight of UBP1 was found to be 83,500 daltons. The optimum temperature and pH for the enzyme reaction when ubiquitin-human growth hormone (hGH) was used as a substrate were $40^{\circ}C$ and pH 8.0, respectively.

• YAKHAK HOEJI
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• v.34 no.6
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• pp.439-446
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• 1990

The general pharmacological actions of recombinant human growth hormone (rHGH) were investigated. It had hypothermic action but neither sedative nor analgesic action. No pharmacological effects were observed in isolated guinea pig ileum and tracheal muscle and rat fundus and uterus. Slight hypotensive action with no effect on respiration was revealed at a dose of 20 IU/kg i.v. of rHGH in rabbits. The rHGH exhibited a weak inhibitory action of glucose tolerance in normal rats, significantly lowered the blood glucose contents in adrenalectomized rats 20 min after i.v. administration (80IU/kg), and produced a significant inhibitory effect on in vitro glycerol release in epinephrine-stimulated epididymal fat pad segments of rats.

• Biomolecules & Therapeutics
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• v.3 no.4
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• pp.294-300
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• 1995

DA-3002, an authentic recombinant human growth hormone(rhGH), was examined for mutagenicity in the reverse mutation test on bacteria, in the chromosomal aberration test on cultured mammalian cells and in the micronucleous test on mice. The reverse mutation test was performed by a plate incorporation method with or without a metabolic activation system(S9 Mix) using Salmonella typhimurium strain TA100, TA1535, TA98 and TA1537. DA-3002 did not significantly increase revertant colonies in any of the test strains under any conditions at dose levels ranging from 0.0125 to 0.4 IU/plate, compared with the vehicle control. In the chromosomal aberration test using cultured Chinese hamster lung(CHL) cells, DA-3002 did not increase the number of aberrant cells in the presence or absence of S9 mix at concentrations of 0.0125 IU/mι to 0.4 IU/mι, compared with the vehicle control. In the micronucleus test, male ICR mice were given DA-3002 intraperitoneally at a dose level of 20, 40 and 80 lU/kg. The incidence of bone marrow micronucleated polychromatic erythrocytes in the DA-3002 treated mice did not differ from that of the vehicle control. These results indicate that DA-3002 doesn't have mutagenic potential under the present test conditions.

• BMB Reports
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• v.34 no.6
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• pp.502-508
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• 2001

A cDNA, encoding the human growth hormone (hGH), was synthesized based on the known 191 amino acid sequence. Its codon usage was optimized for a high level expression in Escherichia coli. Unique restriction sites were incorporated throughout the gene to facilitate mutagenesis in further studies. To minimize an initiation translation problem, a 624-bp cassette that contained a ribosome binding site and a start codon were fused to the hGH-coding sequence that was flanked between the EcoRI and HindIII sites. The whole fragment was synthesized by an overlapped extension of eight long synthetic oligonucleotides. The four-short duplexes of DNA, which were first formed by annealing and filling-in with a Klenow fragment, were assembled to form a complete hGH gene. The hGH was cloned and expressed successfully using a pET17b plasmid that contained the T7 promoter. Recombinant hGH yielded as much as 20% of the total cellular proteins. However, the majority of the protein was in the form of insoluble inclusion bodies. N-terminal amino acid sequencing also showed that the hGH produced in E. coli contained formyl-methionine. This study provides a useful model for synthesis of the gene of interest and production of recombinant proteins in E. coli.

• Proceedings of the Korean Society of Food Hygiene and Safety Conference
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• 1994.12a
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• pp.15-26
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• 1994

Bovine somatotropin(bST) or bovine growth hormone (bGH) is a protein of 191 amino acids produced by the anterior pituitary gland of cattle. Recombinant bovine somatotropin(rbST) is biosynthetic versions of the naturally occurring pituitary hormone in cows. The use of rbST in dairy cows promises to improve the efficiency of milk production around the world. Using recombinant DNA technology, bST can now be produced in commercial quantities. The recombinant bST(rbST) is biologically identical to the found in the bovine pituitary. Milk from rbST-treated cows has been found to have the same nutritional value and composition as milk from untreated cows. In November of 1993, rbST finally was approved by the FDA, nearly 10 years after filing a licence applica-tion. rbST has been one of the most extensively studied animal drug products to be reviewed by the agency. Three scientific facts will help to reassure the public about the safety of the milk suppy.: 1. rbST has no biological activity in humans when indigested orally or when given by intramuscular injection. 2. Insulin-like growth factor 1(IGF-1) is not orally active. Any changes in IGF-1 levels in milk are well within normal variation and are lower than those reported in human milk. 3. All cow's milk contains bST, and no significant change in bST levels in milk occurs as a result of giving cows supplemental bST. Based on the scientific evidence, the public can be confident that milk and meat from rbST-treated cows is safe to consumers.

• Proceedings of the PSK Conference
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• 2003.10b
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• pp.230.3-231
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• 2003

The therapeutic use of protein pharmaceuticals produced by recombinant DNA technology is increasing in recent decades. In order to investigate the quality of recombinant proteins, it is important to identify and assign the impurities produced in the process of recombination or in storage conditions. Capillary Electrophoresis is emerging technology exhibiting high sensitivity, selectivity and speed and may be most powerful tools for this application. In this study, human growth hormone (hGH) has been analyzed by various mode of capillary electrophoresis such as capillary zone electrophoresis (CZE), capillary gel electrophoresis (CGE), and capillary isoelectric focusing (cIEF) to indicate the chemically or biologically oriented variants and the degraded fragments. (omitted)

• Childhood Kidney Diseases
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• v.13 no.1
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• pp.14-20
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• 2009

Growth retardation is a common consequenc of chronic kidney disease (CKD) in childhood. Many recent clinical and experimental data indicate that growth failure in CKD is mainly due to a relative GH insensitivity and functional IGF-I deficiency. Glucocorticoids also glucocorticoids interfere with the integrity of the somatotropic hormone axis at various levels. Over the past 10 years, recombinant growth hormone (rhGH) has been used to help short children with chronic kidney disease. A GH dosage of 0.35 mg/kg/week (28 IU/$m^2$/week) appears efficient and safe. Some clinical trial data show that final height will be within the normal target height range when GH treatment is continued for many years without remarkable adverse events.