• Title/Summary/Keyword: Reproductive Physiology

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Examination of Improved Tetracycline Inducible Gene Expression System In Vitro (새로운 Tetracycline 유도적 유전자 발현 System의 In Vitro 검정)

  • Kwon, Mo Sun;Kim, Teoan;Koo, Bon Chul
    • Reproductive and Developmental Biology
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    • v.37 no.3
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    • pp.109-115
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    • 2013
  • Until recently the most popular tetracycline-inducible gene expression system has been the one developed by Gossen and Bujard. In this study, we tested the latest version of same system and the results are summarized as follows: Compared with previous one, the difference of new system are minor changes of nucleotide sequences in transactivator and tetracycline response element (TRE) regions. Sensitivity to the doxycycline (a tetracycline derivative) was improved. Leakiness of GFP marker gene expression in non-inducible condition was significantly decreased. Higher expression of the marker gene was observed when the cells were fed with doxycycline-containing medium. Optimal insertion site of woodchuck posttranscriptional regulatory element (WPRE) sequence which was known to increase gene expression was different depending on the origin of cells. In chicken embryonic fibroblast, location of WPRE sequence at 3' end of TRE resulted in the highest GFP expression. In bovine embryonic fibroblasts, 3' end of transactivator was the best site for the GFP expression.

Identification of Differentially Expressed Genes between Neonatal and Peripubertal Rat Thymi Using $GeneFishing^{TM}$ Polymerase Chain Reaction

  • Kang, Da-Won;Kim, Gyu-Tae;Han, Jae-Hee
    • Reproductive and Developmental Biology
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    • v.31 no.1
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    • pp.55-60
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    • 2007
  • Aging causes thymus involution, and genes in thymus play an important role in the development of the immune system. In this study, we compared genes expressed in thymus of neonatal and peripubertal rats using annealing control primers (ACPs)-based GeneFishing polymerase chain reaction (PCR) and semiquantitative reverse transcription (RT)-PCR. We identified 10 differentially expressed genes (DEGs) with 20 ACPs. Of 10 DEGs, bystin-like, collagen type V alpha 1 (COL5A1), and T-cell receptor beta-chain segment 2 (TCRB2) that are related to immune-function were detected in rat thymus. Bystin-like and TCRB2 were up-regulated, while COL5A1 was down-regulated in peripubertal thymus. Semiquantitative RT-PCR confirmed postnatal changes in expression of bystin-like, COL5A1, and TCRB2. These results suggest that bystin-like, COL5A1, and TCRB2 could regulate immune function controlled in thymus as age increases.

Generation of Transgenic Chickens Regulating hEPO Gene Expression (hEPO 유전자의 발현이 조절되는 형질전환 닭의 생산)

  • Koo, Bon-Chul;Kwon, Mo-Sun;Kim, Te-Oan
    • Reproductive and Developmental Biology
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    • v.34 no.3
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    • pp.193-199
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    • 2010
  • We report here the production of transgenic chickens that can regulate human erythropoietin (hEPO) gene expression. The glycoprotein hormone hEPO is an essential for viability and growth of the erythrocytic progenitors. Retrovirus vector system used in this study has two features including tetracycline-controllable promoter and woodchuck hepatitis virus posttranscriptional regulator element (WPRE). The former is for to reduce the possibility of physiological disturbance due to constitutional and unregulated expression of hEPO gene in the transgenic chicken. The latter is for maximum expression of the foreign gene when we turn-on the gene expression. A replication-defective Moloney murine leukemia virus (MoMLV)-based vectors packaged with vesicular stomatitis virus G glycoprotein (VSV-G) was injected beneath the blastoderm of non-incubated chicken embryos (stage X). Out of 325 injected eggs, 28 chicks hatched after 21 days of incubation and 16 hatched chicks were found to express the hEPO gene delivered by the vector. The biological activity of the recombinant hEPO in transgenic chicken serum was comparable to its commercially available counterpart. The recombinant hEPO in transgenic chicken serum had N- and O-linked carbohydrate simillar to that produced from in vitro cultured cells transformed with hEPO gene.

Biotechnological Approaches in Sericultural Science and Technology of Uzbekistan

  • Madyarov Shukhrat R.
    • International Journal of Industrial Entomology and Biomaterials
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    • v.11 no.1
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    • pp.13-19
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    • 2005
  • Biotechnological researches in Central Asian Research Institute of Sericulture were developed since 1981 when laboratory of artificial diets for mulberry silk­worm was created. The researches were directed not only to creation of accessible artificial diets (AD) for off-season mass rearing of economical beneficial insect and for industrialization of sericulture but also to obtaining of easy reproducible source of animal proteins, carbohydrates, lipids, enzymes, enzymes inhibitors and other metabolites - bioresource for manufacture of oriental medicine preparations. Space experiments with silkworms became possible with use of AD. These experiments used mulberry silkworm as high effective board test organism with outstanding developmental, reproductive and transgenic properties having impor­tant significance in future of cosmonautics. Introduction to practice of bioprotective and ecologically pure method of drying and sterilizing of green cocoons and by-products of silk manufacture allows to increase efficiency of cocoons reeling technology as well as to preserve nativity of biological active substances in mulberry silkworm pupae that is unachievable by traditional methods of raw material processing. New spheres of application of two important silk proteins - fibroin and sericin in medicine, cosmetics, for bio- and nanotechnology are opening with use of biotechnological approaches. At present the achievements and experience of R & D in these fields are used in creation of modern pests biocontrol agents. Schemes of efficient processing of cocoon raw material, by-products and wastes of sericulture and cocoon processing are devel- Institute of Physiology and Biophysics, Uzbek Academy of Sciences, I, Niyazov street, 700095, Tashkent, Uzbekistan. oped for obtaining of traditional and new consumer products.

In Vitro Assay of Mammary Gland Tissue Specific hEPO Gene Expression (hEPO 유전자의 유선조직 특이적 발현에 대한 In Vitro 검정)

  • Koo, Bon Chul;Kwon, Mo Sun;Kim, Teoan
    • Reproductive and Developmental Biology
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    • v.40 no.1
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    • pp.7-13
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    • 2016
  • Effectiveness of transgene transfer into genome is crucially concerned in mass production of the bio-pharmaceuticals using genetically modified transgenic animals as a bioreactor. Recently, the mammary gland has been considered as a potential bioreactor for the mass production of the bio-pharmaceuticals, which appears to be capable of appropriate post-translational modifications of recombinant proteins. The mammary gland tissue specific vector system may be helpful in solving serious physiological disturbance problems which have been a major obstacle in successful production of transgenic animals. In this study, to minimize physiological disturbance caused by constitutive over-expression of the exogenous gene, we constructed new retrovirus vector system designed for mammary gland-specific expression of the hEPO gene. Using piggyBac vector system, we designed to express hEPO gene under the control of mammary gland tissue specific and lactogenic hormonal inducible goat ${\beta}$-casein or mouse Whey Acidic Protein (mWAP) promoter. Inducible expression of the hEPO gene was confirmed using RT-PCR and ELISA in the mouse mammary gland cells treated with lactogenic hormone. We expect the vector system may optimize production efficiency of transgenic animal and reduce the risk of global expression of transgene.

Proline accumulation and transcriptional regulation of proline biothesynthesis and degradation in Brassica napus

  • Xue, Xingning;Liu, Aihua;Hua, Xuejun
    • BMB Reports
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    • v.42 no.1
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    • pp.28-34
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    • 2009
  • To understand the molecular mechanism underlying proline accumulation in Brassica napus, cDNAs encoding ${\Delta}^1$-pyrroline-5-carboxylate synthetase (BnP5CS), ornithine $\delta$-aminotransferase (BnOAT) and proline dehydrogenase (BnPDH) were isolated and characterized. Southern blot analysis of BnP5CSs in B. napus and its diploid ancestors suggested a gene loss may have occurred during evolution. The expression of BnP5CS1 and BnP5CS2 was induced, while the expression of BnPDH was inhibited under salt stress, ABA treatment and dehydration, prior to proline accumulation. The upregulation of BnOAT expression was only detected during prolonged severe osmotic stress. Our results indicate that stress-induced proline accumulation in B. napus results from the reciprocal action of activated biosynthesis and inhibited proline degradation. Whether the ornithine pathway is activated depends on the severity of stress. During development, proline content was high in reproductive organs and was accompanied by markedly high expression of BnP5CS and BnPDH, suggesting possible roles of proline during flower development.

Clinical Cases Related to Jeong in Hyungsang Medicine (정에 대한 제질환의 형상의학적 임상예)

  • Kim Kyung Chul;Lee Yang Tae
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.16 no.6
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    • pp.1092-1099
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    • 2002
  • Observations are made on the causes and symptoms of diseases related to Jung in 'the precious mirror of oriental medicine' From the study of those clinical cases diagnosed and treated with hyungsang medicine, the following conclusions are drawn Jung is made from shin of parents and it is the basis of the body. it can be derived in to two ; the innate and the acquired. diseases of Jung come from unbalanced meals, immoderate drinking immoderate sexual life, emotional disturbance and irregular daily life. diseases of Jung affect not only reproductive organs but also the body all over. persons with following configurational characteries tic. one easily attacked with diseases Jung ; fish type, bird type, Jung type, shin type, thin, male, pod peoples, pronounced or slanted mouth and nose upturned. nose, congested and inanimated eyes, pale lips, big or red cheekbone, rough skin and grizzled hair. To preserve and satisfy Jung, it is necessary to practice temperance in sexual life, emotions, daily life and meals.

Relationship between Intersequence Pauses, Laying Persistency and Concentration of Prolactin during the Productive Period in White Leghorn Hens

  • Reddy, I.J.;David, C.G.;Singh, Khub
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.5
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    • pp.686-691
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    • 2005
  • Prolactin is considered to influence the taking of pauses in between ovulatory sequences in White Leghorn hens. Therefore modulating concentrations of prolactin using bromocriptine - a dopamine agonist during early life (17 to 36 weeks of age) could overcome the inhibitory effects of high concentration of prolactin on ovarian activity. The effect of modulation of prolactin concentration on egg production, sequence length and inter sequence pauses were studied by analyzing the oviposition records from 19 to 72 weeks were studied and compared with untreated controls. Bromocriptine administered subcutaneously (100 $\mu$g kg$^{-1}$ body weight or orally through feed (640 $\mu$g day$^{-1}$ bird$^{-1}$) resulted in a steady and sustained decrease in prolactin levels (p<0.01) during and after the withdrawal of treatment up to one reproductive cycle (72 weeks of age). The treated birds had comparatively longer sequences (p<0.01) and fewer pauses (p<0.01). Egg production increased (p<0.01) by fourteen per cent through subcutaneous administration and eleven per cent through oral feeding, over the control birds. It is concluded that the physiological pauses that occur during ovulatory sequences can be disrupted effectively using bromocriptine. Prolactin levels are modulated which may interfere with the follicular recruitment and subsequent oviposition thereby improve egg laying potential of the bird.