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토끼 간조직의 Stimulatory Guanine Nucleotide 결합 단백의 αsubunit 에 대한 cDNA 염기서열에 관한 연구
이상훈,노연희,한중수,고재경 한양대학교 의과대학 1993 한양의대 학술지 Vol.13 No.1
G proteins, a family of guanine nucleotide-binding protein superfamily, are involved in transmembrane signaling systems as transducers. In this study cDNAs encoding αsubunit of the stimulatory G protein (G α ) from rabbit liver were cloned and their nucleotide sequences were determined : a λgt 11 cDNA library from total cellular poly (A)' mRNA in rabbit liber was screened for recombinant λDNA that codes for G α with labeled probe. The probe used was an EcoRI digested 3' end fragment of cDNA for G α from olfactory neuroepithelium of rat. Six of the approximately recombinant clones were screened by in situ hybridization and by autoradiography, detecting sixteen positive clones. These clones were tested using polymerase chain reaction (PCR) technique and thirteen of them were turned out to have two sets of conserved sequence in αcDNA. Four clonse among them were selected form analysis of their cDNA sequences, showing the presence of two types of cDNA, namely and with the total length of nucleotide sequence of1.392bp and 1.613bp, respectively . From nucleotide sequence analysis the amino acid residues of -1 and α-2 were deduced : they contain 335 and 386 amino acid residues (including the initiator methionine), respectively. The calculated molecular weights for and were 38.9kd and 44.6kd, respectively. The significant difference in size between two cDNAs for G αseemed to be due to the assumption that alternative promoter and leading exon might be involved in transcribing the mRNA for . Two types of cDNA from rabbit liver shared over 95% of amino acid homology with that from rat olfactory neuroepichelium if deleted portions are not counted. More information on the cDNAs could be obtained through further studies such as Sl nuclease protection experiment, expression and mutation study.
대장균의 GlnE 유전자 조절 부위의 염기서열 분석에 관한 연구
박종환,이영규,노연희,구자현 건국대학교 1996 學術誌 Vol.40 No.2
The enzymatic activity of glutamine synthetase (GS) which is a major enzyme assimilating nitrogen in enteric bacteria is modulated by a bicyclic cascade that consists in adenylylation / deadenylylation of GS and uridylylation / deuridylylation of a regulatory protein PII. The adenylylation of GS converts the enzyme to an inactive form. The cunverter enzyme for the adenylylation / deadenylylation cycle is adenylyltransferase (AT), a bifunctional enzyme. It is known that many of regulatory proteins of GS are affected by the nitrogen content in medium. In this study, glnE gene sequence from BamHI to Sali restriction site was determined to elucidate the possibility of its nitrogen sensing ability by searching the consensus sequence(WGCA) found in other nitrogen utilizing operons. The results are as follows ; 1) The nucleotide length of a part of glnE gene was 1203 dp and the initial codon ATG was determined by comparing the sequence with 13 amino acid residues from N-terminus of AT. 2) The probable consensus sequence TTGCA showing high homology with those of other nitrogen utilizing operons was found at 43 bp upstream region from the start point. Actual nitrogen sensing capability of glnE gene should be tested to find the complex regulatory mechanism of GS cascade system for further study.
Circadian Rhythms in Urinary Functions: Possible Roles of Circadian Clocks?
Noh, Jong-Yun,Han, Dong-Hee,Yoon, Ji-Ae,Kim, Mi-Hee,Kim, Sung-Eun,Ko, Il-Gyu,Kim, Khae-Hawn,Kim, Chang-Ju,Cho, Sehyung Korean Continence Society 2011 International Neurourology Journal Vol.15 No.2
<P>Circadian clocks are the endogenous oscillators that harmonize a variety of physiological processes within the body. Although many urinary functions exhibit clear daily or circadian variation in diurnal humans and nocturnal rodents, the precise mechanisms of these variations are as yet unclear. In this review, we briefly introduce circadian clocks and their organization in mammals. We then summarize known daily or circadian variations in urinary function. Importantly, recent findings by others as well as results obtained by us suggest an active role of circadian clock genes in various urinary functions. Finally, we discuss possible research avenues for the circadian control of urinary function.</P>
Noh, Yun-Hee 건국대학교 의과학연구소 2002 건국의과학학술지 Vol.12 No.-
Thrombospondin-2(TSP-2) has been known to modulate tumor growth, wound healing, and inflammation through inhibition of angiogenesis based on the findings from TSP-2 knock-out mice and overexpression studies. However, the molecular mechanisms of human TSP-2(hTSP-2) for inhibition of angiogenesis or the feasibility to develope as a new therapeutic angiogenesis inhibitor for curing cancer have not been intensively studied because so far the whole molecule of hTSP-2 is not available due to difficulties in extraction of useful amount of hTSP-2 from tissues and limitations on production of full length recombinant hTSP-2 in mammalian or insect cells. Therefore, an N-terminal 80 kDa fragment of hTSP-2 encompassing N-terminal globular region through typeⅠ repeats(hTSP-2/NTF) was expressed in human embryonal kidney 293 EBNA cells to investigate whether this fragment has antiangiogenic effect on primary cultured human endothelial cells(ECs). The recombinant vector was constructed as follows: an 1.67 kbp of cDNA of hTSP-2(nt 213 to 1883) was amplified by PCR from human placenta cDNA library and ligated into KpnⅠ and Bgl Ⅱ sites of the modified pCEP 4 expression vector. The recombinant protein was purified from the serum fee conditioned medium of transfected 293 EBNA cells using ammonium sulfate precipitation and gelatin- and heparin-sepharose columns. Yield was 2.1 ㎎/l of conditioned medium and the purified protein was confirmed by N-terminal peptide sequencing. To see the fragment has anti-angiogenic activity in vitro, migration assay was performed on human dermal microvascular ECs(HDMECs) with the recombinant protein, demonstrating inhibition of migration by 30 % at 1 ㎎/ml of hTSP-2/NTF as compared to control group. This effect was not neutralized by anti-CD 36 antibody, suggesting a different receptor other than CD 36, to which TSP-1 binds and inhibits EC migration, may be involved in the inhibitory mechanism of hTSP-2/NTF on the migration of HDMECs. Futher study to see in vivo antiangiogenic activity of hTSP-2/NTF is on-going to evaluate the validity of this fragment as a new antiangiogenic agent.
Noh, Jong-Yun,Han, Dong-Hee,Kim, Mi-Hee,Ko, Il-Gyu,Kim, Sung-Eun,Park, Noheon,Kyoung Choe, Han,Kim, Khae-Hawn,Kim, Kyungjin,Kim, Chang-Ju,Cho, Sehyung Nature Publishing Group 2014 Experimental and molecular medicine Vol.46 No.3
<P>Circadian clocks are the endogenous oscillators that harmonize a variety of physiological processes within the body. Although many urinary functions exhibit clear daily or circadian variation in diurnal humans and nocturnal rodents, the precise mechanisms of these variations are as yet unclear. In the present study, we demonstrate that <I>Per2</I> promoter activity clearly oscillates in neonate and adult bladders cultured <I>ex vivo</I> from <I>Per2</I>::Luc knock-in mice. In subsequent experiments, we show that multiple local oscillators are operating in all the bladder tissues (detrusor, sphincter and urothelim) and the lumbar spinal cord (L4–5) but not in the pontine micturition center or the ventrolateral periaqueductal gray of the brain. Accordingly, the water intake and urine volume exhibited daily and circadian variations in young adult wild-type mice but not in <I>Per1</I><SUP>−/−</SUP><I>Per2</I><SUP>−/−</SUP> mice, suggesting a functional clock-dependent nature of the micturition rhythm. Particularly in PDK mice, the water intake and urinary excretion displayed an arrhythmic pattern under constant darkness, and the amount of water consumed and excreted significantly increased compared with those of WT mice. These results suggest that local circadian clocks reside in three types of bladder tissue and the lumbar spinal cord and may have important roles in the circadian control of micturition function.</P>