Circadian Expression of Cryptochrome in Rat Brain and Muscle

Kang, Hae Mook 청주대학교 산업과학연구소 2006 産業科學硏究 Vol.23 No.2

포유류의 일주기 조절에는 두 종류의 cryptochrome(Cry1과 Cry2)이 관여하는 것으로 알려져 있다. 흰쥐에서 Cry1과 Cry2 유전자의 일주기적인 발현을 suprachiasmatic nucleus(SCN)조직과 근육 조직에서 real-time PCR로 조사하였다. SCN 조직에서 Cry1과 Cry2의 발현 정도는 밤낮의 변화에 따른 일주기적인 리듬을 보여주고 있다. 근육 조직 또한 SCN과는 약간 다르지만 분명하게 Cry1과 Cry2의 일주기적인 발현을 하고 있다. 따라서 이 결과는 Cry1과 Cry2가 SCN 조직에서 중추적인 생체시계와 근육조직에서 말단 생체시계의 역할을 하고 있음을 강력히 시사하고 있다. In mammals, two types of cryptochrome (Cry1 and Cry2) are known to involve in the regulation of circadian rhythm. Circadian expression of rat Cry1 and Cry2 mRNA was examined in suprachiasmatic nucleus (SCN) and muscle tissues by real-time PCR. The expression level of Cry1 and Cry2 mRNA in SCN showed circadian rhythm with a peak at the time of day/night transition. Muscle also exhibited the circadian pattern of the expression of Cry1 and Cry2 mRNA at slightly different with that of SCN. It is, therefore, strongly suggested that Cry1 and Cry2 play an important role in central clock of SCN and peripheral clock in muscle.

슬럼프값을 갖지 않는 콘크리트에 관한 연구

鄭海駿,姜準默,李鎭沃 충남대학교 공업교육연구소 1982 論文集 Vol.5 No.1

This paper identified the W/C ratio of no-slump dr lean concrete analyses its compressive strength and its coefficient of permeability. The relation between no-slump W/C ratio and the quantity of unit cement is approximately linear. The compressive strength by variation of W/C was relatively low and the coefficient of permeability was comparatively high. By the use of admixture, a better quality of no-slump dry lean concrete is expected.

Wire Pieces를 添加한 Concrete Beam의 휨引張 强度에 關한 硏究

鄭海駿,姜準默 충남대학교 공업기술개발연구소 1975 工業技術開發硏究所論文集 Vol.2 No.2

Todays the range that concrete material occupies of all the constructive materials is larger than any other material. Concrete material occupying the major part as all the constructive materials has a lot of merits, but the greatest defect is that tensile strength is low to compressive strength as we can disregard its values in engineering. Therefore concrete needs a certain material reinforcing this. As reinforce, we have reinforced the defect of concrete by inserting metal rinforcement in it. In this study, thinking a great many wire pieces can be produced, dynamic character is excellent, and construction method is easy, the authors used wire pieces as a mixture to reinforce concrete. This experimental result is to present a practical possibility.

高張力 Bolt를 Dowel로 利用한 合成桁에 關한 硏究

鄭海駿,姜準默 충남대학교 공업기술개발연구소 1976 工業技術開發硏究所論文集 Vol.3 No.2

This paper is a research on the coefficient of friction occured between the channel and concrete through a push-out testing. For preparing the test pieces, the authors meaured the size of a concrete slab according to the specification of the composite girder design, using the high tension bolts instead of various dowels for shear connector. The result showed the possibility to design composite girder under the condition of the coefficient of friction μ=0.38 using the high tension bolts.

Session F : Eukaryotic Gene Structure and Expression : Production and analysis of transgenic mice carrying phospholipase C gene disrupted by gene targeting . Ⅰ . Cloning of the PLC - β1 locus and construction of the targeting vector

Hae Mook Kang,Dae Soo Kim,Nae Kyu Kang,Hee Sup Shin 생화학분자생물학회(구 한국생화학분자생물학회) 1993 추계학술대회집 Vol.- No.-

Molecular Characterization of Amphibian Cryptochrome cDNA as A Clock Gene

Kang, Hae Mook 청주대학교 산업과학연구소 2002 産業科學硏究 Vol.20 No.1

Cryptochrome은 DNA photolyase/flavoprotein 중 하나로 최근 사람을 포함하는 모든 동식물의 24시간 생체주기를 조절하는 광수용체로 알려졌다. 본 논문에서는 양서류에서 Cryptochrome(CRY) cDNA를 클로닝하여 특성을 분석하였다. 먼저 황소개구리의 뇌에서 RNA를 추출한 후 역전사 효소를 사용하여 cDNA를 합성한 후 Cryptochrome 유전자의 degernerate primer들을 사용하여 442 bp크기의 cry2 cDNA 단편을 증폭시켰다. 이 cDNA 단편을 probe로 사용하여 황소개구리 뇌의 cDNA library screening을 한 결과 완전한 크기의 Rana cry2 cDNA를 분리하였다. 클론된 Rana cry2는 1916bp로 염기서열에 기초한 추정된 아미노산 서열은 534개였다. 다른 종의 CRY 단백질과 비교한 결과 약 70% 이상의 높은 동일성을 보여주었다. 따라서 양서류의 cry2 유전자는 성공적으로 클로닝 되었음을 알 수가 있었고, 앞으로 개구리와 같은 야생동물의 24시간 생체주기와 관련된 생리적인 특성을 규명하는 데 이 유전자는 매우 유용할 것으로 사료된다. Cryptochromes (CRY), members of the DNA photolyase/cryptochrome flavoprotein family, regulate the circadian clock in animals and plants. Recently, two cryptochromes, called CRY1 and CRY2, have been isolated in human and mouse. In this paper, to study circadian rhythm in amphibian, cry cDNA in bullfrog (Rana catesbeiana) has been identified and characterized. Firstly, a size of 1916 bp cDNA of Rana cry2 were isolated from bullfrog's brain cDNA library by screening with a partial cry cDNA probe of 442 bp, which obtained by PCR amplification with degenerate primers. The cloned bullfrog cryptochrome cDNA has a complete single open reading frame composed of 543 amino acid. The Blast-search of the deduced amino acid sequence exhibited high homology more than 70 % in human, mouse, and zebrafish CRY2. Taken together, Rana cry2 cDNA has been successfully cloned and it will be very helpful to study the clock genes and the circadian rhythm in wild animal.

Phosphorylation Patterns of the Hypothalamic Proteins : Effects of Forskolin , Phorbol Ester and Ovarian Steroids

Kang, Hae Mook,Kim, Kyung Jin,Cho, Wan Kyoo,Lee, Byung Ju,Kang, Sang Soo,Cho, Byung Nam 한국유전학회 1989 Genes & Genomics Vol.11 No.4

Present study aims to clarify the phosphorylation patterns of proteins derived from the rat hypothalamus pretreated with forskolin and phorbol ester in vitro or ovarian steroids in vivo. Proteins were employed to cell free phosphorylation with γ-^(32) P-ATP, analysed by SDS-polyacryl-amide gel electrophoresis and autoradiography. Treatments of forskolin and phorbol ester resulted in intensifying the specific phosphorylation bands of 43 Kd and 130 Kd, respectively, In vivo treatment of ovarian teroids such as estrogen and/or progesterone were highly effective in modifying the phosphorylation patterns of hypothalamic proteins. Furthermore, changes in protein phosphorylation by forskolin, phorbol ester and ovarian steroids were very similar to those with kinase activators such as cAMP, Ca^(2+)-phosphatidylserine and Ca^(2+)-calmodulin. Therefore, it appears that protein phosphorylation is an important step in regulating the trans-synaptic signal transduction in the hypothalamus.

Cloning of cyclin cDNA involved in control of cell cycle in amphibian

Kang, Hae Mook 청주대학교 산업과학연구소 1998 産業科學硏究 Vol.16 No.-

세포주기 연구를 위한 한 모델로에서 양서류 난자성숙과정은 많이 얀구되어 왔다. 본 연구는 독특한 난자성숙 기작을 보여주는 북방산 개구리 (Rana dybowskii) 난자에서 MPF의 분자적 특성을 밝히고자, 먼저 MPF의 한 성분인 cyclin B1 유전자를 클로닝 하였다. 이미 밝혀진 척추동물의 cyclin B 유전자의 보존된 염기서열에 기초하여 올리고염기를 제작하여 RT-PCR를 북방산 개구리의 개놈 DNA 라이버리에서 행하였다. 그 결과 약 300bp의 DNA 조각을 얻었고, 이의 염기서열을 결정하고 분석한 결과 사이클린 B1 유전자임 밝혀졌다. Northern 분석 결과 cyclin B1 유전자의 전사크기는 약 1.6 kb인 것으로 추정되고, 주로 난소나 정소와 같은 생식소에서 발현되는 것을 알수가 있었다. 이상의 연구 결과 북방산 개구리의 난자에 MPF가 존재함을 추정할 수가 있고, 이는 계절적인 번식을 하는 동물의 세포주기를 밝힐 수 있는 분자적 단서를 제공하는 것으로 사료된다. To understand the mechanism of meiotic maturation as a model of cell cycle study, it needs the characterization of MPF and regulation mechanism of MPF activation in R. dybowskii. To clone a cyclin cDNA from R. dybowskii ovary cDNA library, polymerase-chain reaction (PCR) was performed with degenerate oligonucleotides based on the conserved sequence of vertebrate cyclin B genes. As a result of PCR, about 300 bp DNA fragment was amplified Sequencing and Blast search indicated that it encodes of cyclin Bl gene of Rana. Northern blot showed that a single band of 1.6 kb was detected in both of ovary and testis tissues. It, therefore, indicate that a partial fragment of cyclin Bl cDNA was cloned, and mainly expressed in gonadal tissues in Rana dybowskii. It Is also feasible to reveal the character and molecular structure of MPF involved control of cell cycle in seasonal breeding amphibian.

Dicistronic Tagging of Genes Active in Embryonic Stem Cells of Mice

Kang, Hae Mook,Kang, Nae-Gyu,im, Dae-Gohn,shin, Hee-Sup Korean Society of Molecular Biology 1997 Molecules and cells Vol.7 No.4

Comparative Study on the Molecular Evolution of GnRH Gene in Amphibia

Kang, Hae Mook 청주대학교 산업과학연구소 2000 産業科學硏究 Vol.18 No.1

양서류에서 GnRH 유전자의 분자적 진화과정을 연구하기 위해, 북방산 개구리 (Rana dybowsku)와 황소개구리 (Rana catesbeiana)에서 GnRH 전구체의 유전자를 클로닝하였다. 북방산 개구리의 GnRH cDNA는 90개의 아미노산을 가진 약 500 bp였다. 추정된 GnRH 분자는 8번째 아미노산이 tryptophane인 새로운 형태의 GnRH였다. 황소개구리의 GnRH cDNA는 약 648 bp로 구성되어 있으며, 270 bp로 된 하나의 open-reading frame을 가지고 있다. 추정된 황소개구리의 GnRH 분자는 포유류의 GnRH와 동일하였다. 이들 개구리의 GnRH 전구체들은 서로 높은 동질성을 보여주고 있으며, 반면에 다른 척추동물의 GnRH 분자와의 동질성은 낮았다. 따라서 양서류 GnRH는 어류에서 진화되었음을 알 수가 있었다. To study the molecular evolution of GnRH cDNA in amphibian, cDNAs encoding the GnRH precursors were isolated from Korean name frog (Ram dybowsku) and bullfrog (Rana catesbeiana). The GnRH cDNA of R dybowsku consists of 506 bp and contains an open-reading frame of 270 nucleotides, encoding the GnRH precursor. The GnRH gene encodes a GnRH peptide ([Trp8]GnRH) in which tryptophan is substituted for arginine of mammalian GnRH. The bulfrog GnRH cDNA consists of 648 bp and contains an open-reading frame of 270 nucleotides, encoding the GnRH precursor Both types of frog GnRH precursor have a similar molecular architecture as observed in other GnRH precursors, consisting of a signal peptide, followed by the GnRH decapeptide, a conserved carboxy-terminal amidation and proteolytical processing site, and a GnRH-associated peptide (GAP). Molecular phylogenic analysis reveals that the cloned frog GnRH precusors clearly belongs to the hyptothalamic GnRH Frog GnRH precusors shows a relatively high homology among them, but a less homology with other vertebrates.