• Asian-Australasian Journal of Animal Sciences
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• 제20권7호
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• pp.1023-1029
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• 2007

Inhibins participate in the regulation of pituitary follicle-stimulating hormone synthesis and secretion, follicular maturation and steroidogenesis in the female. Inhibin ${\beta}_A$ gene (INHBA) was studied as a candidate gene for the prolificacy of sheep. Single nucleotide polymorphisms of the entire coding region and partial 3' untranslated region of INHBA were detected by PCR-SSCP in two high fecundity breeds (Small Tail Han and Hu sheep) and six low fecundity breeds (Dorset, Texel, German Mutton Merino, South African Mutton Merino, Chinese Merino and Corriedale sheep). Only the PCR products amplified by primers 3, 4 and 5 displayed polymorphisms. For primer 3, genotype CC was only detected in Chinese Merino sheep, genotype AA was detected in the other seven sheep breeds. Genotype BB was only detected in Hu sheep. Only Hu sheep displayed polymorphism. Eight or four nucleotide mutations were revealed between BB or CC and AA, respectively, and these mutations did not result in any amino acid change. For primer 4, genotypes EE, EG and GG were detected in Dorset and German Mutton Merino sheep, genotypes EE, EF and FF were detected in Chinese Merino sheep, only genotype EE was detected in the other five sheep breeds. Only Dorset, German Mutton Merino and Chinese Merino sheep displayed polymorphism. Sequencing revealed one nucleotide mutation ($114G{\rightarrow}A$) of exon 2 of INHBA gene between genotype FF and genotype EE, and this mutation did not cause any amino acid change. Another nucleotide change ($143C{\rightarrow}T$) was identified between genotype GG and genotype EE, and this mutation resulted in an amino acid change of $serine{\rightarrow}leucine$. For primer 5, genotypes KK and KL were detected in German Mutton Merino and Corriedale sheep, genotypes KK, LL and KL were detected in the other six sheep breeds. Genotype MM was only detected in Hu sheep. All of these eight sheep breeds displayed polymorphism. Sequencing revealed one nucleotide mutation ($218A{\rightarrow}G$) of exon 2 of the INHBA gene between genotype LL and genotype KK, and nine nucleotide mutations between genotype MM and genotype KK. These mutations did not alter amino acid sequence. The partial sequence (395 bp for exon 1 and 933 bp for exon 2) of the INHBA gene in Small Tail Han sheep (with genotype KK for primer 5) was submitted into GenBank (accession number EF192431). Small Tail Han sheep displayed polymorphisms only in the fragment amplified by primer 5. The Small Tail Han ewes with genotype LL had 0.53 (p<0.05) or 0.63 (p<0.05) more lambs than those with genotype KL or KK, respectively. The Small Tail Han ewes with genotype KL had 0.10 (p>0.05) more lambs than those with genotype KK.

• Bulletin of the Korean Chemical Society
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• 제26권8호
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• pp.1289-1292
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• 2005

• Journal of Pharmaceutical Investigation
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• 제25권3호spc1호
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• pp.7-20
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• 1995

Taurine, a ${\beta}-amino$ acid, plays an important role as a neuromodulator and is necessary for the normal development of the brain. Since de novo synthesis of taurine in the brain is minimal and in vivo studies suggest that taurine dose not cross the blood-brain barrier, we examined whether the choroid plexus, the blood-cerebrospinal fluid (CSF) barrier, plays a role in taurine transport in the central nervous system. The uptake of $[^3H]-taurine$ into ATP depleted choroid plexus from rabbit was substantially greater in the presence of an inwardly directed $Na^+$ gradient taurine accumulation was negligible. A transient in side-negative potential gradient enhanced the $Na^+-driven$ uptake of taurine into the tissue slices, suggesting that the transport process is electrogenic, $Na^+-driven$ taurine uptake was saturable with an estimated $V_{max}$ of $111\;{\pm}\;20.2\;nmole/g/15\;min$ and a $K_M\;of\;99.8{\pm}29.9\;{\mu}M$. The estimated coupling ratio of $Na^+$ and taurine was $1.80\;{\pm}\;0.122.$ $Na^+-dependent$ taurine uptake was significantly inhibited by ${\beta}-amino$ acids, but not by ${\alpha}-amino$ acids, indicating that the transporter is selective for ${\beta}-amino$ acids. Since it is known that the physiological concentration of taurine in the CSF is lower than that in the plasma, the active transport system we characterized may face the brush border (i.e., CSF facing) side of the choroid plexus and actively transport taurine out of the CSF. Therefore, we examined in vivo elimination of taurine from the CSF in the rat to determine whether elimination kinetics of taurine from the CSF is consistent with the in vitro study. Using a stereotaxic device, cannulaes were placed into the lateral ventricle and the cisterna magna of the rat. Radio-labelled taurine and inulin (a marker of CSF flow) were injected into the lateral ventricle, and the concentrations of the labelled compounds in the CSF were monitored for upto 3 hrs in the cisterna magna. The apparent clearance of taurine from CSF was greater than the estimated CSF flow (p<0.005) indicating that there is a clearance process in addition to the CSF flow. Taurine distribution into the choroid plexus was at least 10 fold higher than that found in other brain areas (e. g., cerebellum, olfactory bulb and cortex). When unlabelled taurine was co-administered with radio-labelled taurine, the apparent clearance of taurine was reduced (p<0.0l), suggesting a saturable disposition of taurine from CSF. Distribution of taurine into the choroid plexus, cerebellum, olfactory bulb and cortex was similarly diminished, indicating that the saturable uptake of taurine into these tissues is responsible for the non-linear disposition. A pharmacokinetic model involving first order elimination and saturable distribution described these data adequately. The Michaelis-Menten rate constant estimated from in vivo elimination study is similar to that obtained in the in vitro uptake experiment. Collectively, our results demonstrate that taurine is transported in the choroid plexus via a $Na^+-dependent,saturable$ and apparently ${\beta}-amino$ acid selective mechanism. This process may be functionally relevant to taurine homeostasis in the brain.

• 한국미생물·생명공학회지
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• 제7권4호
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• pp.217-223
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• 1979

자연계에서 분리한 곰팡이 중 전분을 잘 당화하며 산생성력이 강한 한 균주를 선별하여 현재 막걸리 제국용 균종으로 많이 사용되고 있는 백국균(Aspergillus Kawachii)과 비교하여 각종 성분을 경시적으르 분석한 결과는 다음과 같다. 1) 분리균중 당화력 및 액화력이 비교적 강한 M-80을 공시균으로 선별하였다. 2) 백국균과 분리균의 $\alpha$-amylase 역가는 140W. V.로 비슷하였으나 $\beta$-amylase 역가는 54 A. U. 로 분리균이 높았다. 3) acid protease 역가는 분리균이, alkaline pro-tease 역가는 백국균이 높았다. 4) 총산, ethanol, fusel oil의 생성량은 분리균이 약간 높았으며 methanol 함량은 비슷하였다. 5) 술덧중의 유리 아미노산의 함량은 분리균이 백국균보다 약 10%정도 높았으며, 15종의 유리아미노산이 확인되었다.

• Journal of Microbiology and Biotechnology
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• 제8권3호
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• pp.214-221
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• 1998

A Brevibacterium ammoniagenes gene coding for glucose/mannose-specific enzyme II ($EII^{Glc}$) of the phosphoenolpyruvate-dependent phosphotransferase system (PTS) was cloned by complementing an Escherichia coli mutation affecting a ptsG gene, and the complete DNA nucleotide sequence was determined. The cloned gene was identified to be a ptsG, which enables the E. coli transportment to use glucose more efficiently than mannose as the sole carbon source in an M9 minimal medium. The ptsG gene of B. ammoniagenes consists of an open reading frame of 1,983 nucleotides putatively encoding a polypeptide of 661 amino acid residues and a TAA stop codon. The deduced amino acid sequence of the B. ammoniagenes $EII^{Glc}$ shows, at $46\%$, the highest degree of sequence similarity with the Corynebacterium glutamicum EII specific for both glucose and mannose. In addition, the $EII^{Glc}$ shares approximately $30\%$ sequence similarities with sucrose-specific and ${\beta}$-glucoside-specific EIIs of the several bacteria belonging to the glucose-PTS class. The 161-amino-acid C-terminal sequence of $EII^{Glc}$ is also similar to that of E. coli enzyme $IIA^{Glc}$, specific for glucose ($EIIA^{Glc}$). The B. ammoniagenes $EII^{Glc}$ consists of three domains; a hydrophobic region (EIIC) and two hydrophilic regions (EIIA, EIIB). The arrangement of structural domains, IIBCA, of the $EII^{Glc}$ is identical to those of EIIs specific for sucrose or ${\beta}$-glucoside. While the domain IIA was removed from the B. ammoniagenes $EII^{Glc}$ the remaining domains IIBC were found to restore the glucose and mannose-utilizing capacity of E. coli mutant lacking $EII^{Glc}$ activity with $EIIA^{Glc}$ of the E. coli mutant. $EII^{Glc}$ contains a histidine residue and a cysteine residue which are putative phosphorylation sites for the protein.

• 한국작물학회지
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• 제39권3호
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• pp.278-284
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• 1994

콩단백질의 황 아미노산함량은 가축 영양학상 중요한 위치를 차지하기 때문에 신계통이 가져야만 할 필수조건일지도 모른다. 콩 계통간에 저장단백질의 유전적변이가 존재한다면 이는 기존의 육종방법을 통하여 콩의 종자단백질 구성성분을 유전적으로 변경하여 품질을 개량할 수 있는 가능성을 시사하고 있다. 본 연구는 여러 문헌에 보고된 콩종자 저장단백질의 돌연변이 계통들을 선별하여 콩단백질의 품질을 향상시키기 위한 육종 재료로서의 가능성을 평가하기 위하여 실행되었다. 수집된 돌연변이 계통들은 저장단백질의 또 다른 특성을 나타내었다. 그 돌연변이 계통들 중에서 Keburi(P.I.417016), Keburi(P.I.506817), P.I.154608-1 등은 황 아미노산 함량이 상대적으로 다른 돌연변이 계통보다 높은 1.9, 2.1, 1.8%를 나타내었으며, 이는 7S 단백질인 ${\alpha}$ ', ${\alpha}$ , ${\beta}$단백질 함량이 상대적으로 낮기 때문인 것으로 나타났다. 그러므로 그 돌연변이 계통들 중에서 Keburi(P.I.417016), Keburi(P.I.506817), P.I.54608-1 등은 황 아미노산 함량을 향상시키기 위한 중요한 육종재료로, 그 외 돌연변이 계통들은 다른 용도의 육종 재료로 이용할 수 있을 것으로 추측된다.

• Journal of Microbiology and Biotechnology
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• 제11권4호
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• pp.685-692
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• 2001

A gene, Egl, from Paenibacillus sp. KCTC 8848P encoding endo-${\circ}$-1,4-glucanase was cloned and expressed in Escherichia coli. It consisted of an open reading frame of 1,191 bp for a protein that consisted of 397 amino acids with a molecular weight of 44,539 Da. The deduced amino acid sequence of the endo-${\circ}$-1,4-glucanase gene had a 94% similarity to the endo-$\beta$-1,4-glucanase of Bacillus polymyxa. The Egl gene was also expressed in Saccharomyces cerevisiae secreting Endomyces fibuliger $\beta$-glucosidase (BGL1) under the control of the alcohol dehydrogenase (ADC1) gene promoter, S. cerevisiae transformant producing both endo-${\circ}$-1,4-glucanase and ${\circ}$-glucosidase grew on carboxymethyl cellulose as the sole carbon source.

• Applied Biological Chemistry
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• 제51권4호
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• pp.299-304
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• 2008

A ${\beta}$-1,4-endoglucanase gene from Bacillus subtilis H12 was cloned into Escherichia coli JM109 (pBC8) and sequenced. The endoglucanase gene with an insert DNA of 2.5 kb possessed an open reading frame of 1,500 bp encoding a mature protein of 499 amino acids with a calculated molecular mass of 55 kDa. The deduced amino acid sequence showed similarity to those of the known neutral cellulase genes of B. subtilis PAP115 (99.2%) and BSE616 (97.8%), as well as the alkaline gene of Bacillus sp. N4 (55.1%). The endoglucanase activity expressed by E. coli (pBC8) was localized in the periplasmic fraction (80%) and the cytoplasmic fraction (20%). An endoglucanase was purified from the periplasmic fraction by performing gel filtration and anion exchange chromatography. The molecular weight of the purified enzyme was estimated to be 31 kDa by SDS-PAGE, and the maximum activity occurred at pH 7 and $40^{\circ}C$. The enzyme easily hydrolyzed soluble substrates such as carboxymethyl cellulose and barely ${\beta}$-glucan, whereas the sigmacell and xylan, the known insoluble substrates, were not entirely hydrolyzed.

• 한국미생물·생명공학회지
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• 제25권2호
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• pp.137-143
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• 1997

Porcine transforming growth factor-$\beta$1 (TGF-$\beta$1) was expressed in Escherichia coli using cDNA of TGF-$\beta$1 and glutathione S-transferase (GST) fusion vector pGEX-1$\lambda$T. An ApoI-Tth111I fragment of cDNA which correspond to the amino acid residues from 123 to 390 of the precursor TGF-$\beta$1 was inserted into EcoRI-Tth111I digested pGEM#-l$\lambda$T and the recombined plasmid was named pGET-12. Gene products from the cloned regions of the recombinant plasmids pGET-12 was not detected in soluble fraction of cell free extract but detected in insoluble fraction. The solubilization of insoluble gene product was achieved by the treatment of N-laurylsarcosine. Molecular weight of partially purified proteins determined by electrophoresis was same as expected from cloned fragment. The ELISA test results of the purified proteins showed that immunologically detectable epitope was preserved in recombinant protein.

• 한국식품과학회지
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• 제31권6호
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• pp.1458-1464
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• 1999

우리 나라 연근해에서 어획되고 있는 멸치, 밴댕이 및 까나리의 맛 성분조성을 밝히기 위하여 수산물에 널리 분포하고 있는 함질소 엑스성분인 유리아미노산, combined amino acid류, ATP관련 화합물, 4급암모늄 염기 및 구아니시노 화합물 등을 분석하여 상호 비교하였다. 엑스분 질소함량은 밴댕이가 633 mg/100 g으로서 가장 높고, 다음은 멸치로서 601 mg이었으며 까나리는 455 mg으로서 가장 낮았다. 어류 3종에서 $31{\sim}32$종의 다양한 유리아미노산이 검출되었으나, 그 조성은 모두 유사한 경향이었고, 함량이 풍부한 것들로서는 histidine, taurine, alanine, leucine, carnosine, glutamic acid, lysine 등이었다. 그러나 그 함량에는 어종별로 차이가 있어서 밴댕이, 멸치, 까나리에서 각각 1,672 mg/ l00g, 1,416 mg 및 951 mg이었다. Combined amino acid류 함량을 유리아미노산 수준과 비교하면 까나리에서 110%, 멸치에서 84.8%, 그리고 밴댕이에서는 62.4%로서 어종별로 차이가 있었다. ATP관련 화합물은 밴댕이, 멸치, 까나리에서 각각 $6.15{\mu}mol/g,\;4.14{\mu}mol,\;3.81{\mu}mol$로서 어종에 따른 차이가 많았다. Betaine류는 멸치와 까나리에서 glycinebetaine, ${\beta}-alaninebetaine,\;{\gamma}-butyrobetaine$이 그리고, 밴댕이에서는 ${\beta}-alaninebetaine$이 검출되었으나 미량에 불과하였다. TMAO는 까나리에서 265 mg, 멸치 249 mg, 그리고 밴댕이에서는 201 mg이었으며, TMA는 모두 12 mg 이하로서 비슷하였다. Creatine 함량은 밴댕이 242 mg, 멸치 146 mg, 까나리 131 mg으로서 차이가 있었다. 엑스분 중의 질소 분포는 유리아미노산과 combined amino acid류 질소가 가장 높아서 멸치, 밴댕이, 까나리에서 각각 71.1%, 69.0% 및 68.7%로서 유사하였으며, 엑스분 질소의 회수율은 각각 90.8%, 93.2%, 95.0%이었다.