• Journal of Photoscience
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• 제9권2호
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• pp.269-271
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• 2002

To investigate the visual and extra-ocular photoreception, we cloned the opsin genes in ayu (Plecoglossus allivelis). Amplified fragments encoding exon-4 (-5) of opsin cDNAs were cloned from the retina and brains of ayu, and sequenced. One clone was identified as rod (AYU-Rh), two as green cone (AYU-GI, -G2), one as red cone (A YU-R), two as ultraviolet cone (AYU-UVl, UV2), one as VA (AYU-VA), and one as extra-ocular rod (AYU-ExoRh) opsins. 335 amino acids sequence deduced from the full-length cDNA of AYU-Rh showed high identity with that of other fish. Southern blotting analysis indicated that ayu possess two 'rhodopsin' genes, one is visual rhodopsin and the other is non-visual extra-ocular rhodopsin. In situ hybridization showed that the mRNA of AYU-Rh was localized only in rod cells in the retina. On the other hands, AYU-ExoRh was expressed only in the pineal. We cloned two isoforms (AYU-VAM and -VAL) of VA opsin from ayu. The deduced amino acid sequences of these variants were identical to each other within the first 342 residues, but they showed divergence in the C-terminal sequence. AYU- VAL corresponded to the long isoform found in other fish, and AYU-VAM was identified as a new type of VA opsin variant. Pal-VAM is a new probably functional non-visual photoreceptive molecule in fish.

• Ocean Science Journal
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• 제40권1호
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• pp.55-59
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• 2005

We cloned the complete cDNA of the ${\beta}-bubulin$ from the soft coral, Scleronephthya gracillimum $(K\ddot{u}kenthal)$ (Alcyonacea, Octocorallia, Anthozoa, Cnidaria), via the random sequencing of a cDNA library and the 5'-rapid amplification of cDNA end (RACE) technique. The full-length cDNA of the S. gracillimum ${\beta}-tubulin$ comprised 1541 bp, not including the poly $A^+$ stretch, also contained a complete open reading frame, which codes for a total of 445 amino acids. The amino acid residues 16402 appeared to be in a state of conservation in a variety of animals. Northern blot analysis clearly demonstrated that the sequence we have obtained is, indeed, the full-length cDNA of the ${\beta}-bubulin$ gene in S. gracillimum.

• Molecules and Cells
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• 제25권1호
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• pp.142-147
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• 2008

We recently used degenerate PCR and locus-specific PCR methods to identify the endogenous retroviruses (ERV) in the bovine genome. Using the ovine ERV classification system, the bovine ERVs (BERVs) could be classified into four families. Here, we searched the most recently released bovine genome database with the partial nucleotide sequence of the pro/pol region of the BERV ${\beta}3$ family. This allowed us to obtain and analyze the complete genome of BERV ${\beta}3$. The BERV ${\beta}3$ genome is 7666 nucleotides long and has the typical retroviral organization, namely, 5'-long terminal repeat (LTR)-gag-pro-pol-env-LTR-3'. The deduced open reading frames for gag, pro, pol and env of BERV ${\beta}3$ en- code 507, 271, 879 and 603 amino acids, respectively. BERV ${\beta}3$ showed little amino acid similarity to other betaretroviruses. Phylogenetic analysis showed that it clusters with HERV-K. This is the first report describing the genetic structure and sequence of an entire BERV.

• 한국미생물·생명공학회지
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• 제22권1호
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• pp.31-36
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• 1994

From the pSKH201 plasmid which had been previously cloned in our laboratory, a 3.0kbp insert DNA encoding the alkaline phosphatase of Kluyveromyces fragilis was cleaved with several restriction endonucleases and ligated int the appropriate sites of M13mp18/19 vectors and sequenced by Sanger's dideoxy chain termination method. The sequence contained a 1,638 bp open reading frame(ORP) whose similarities in nucleotide, when compared with those of Saccharomyces cerevisiae and Escherichia coli by GENETYX program, were found to be 61% and 46%, respectively. The deduced amino acid sequence consists of 546 amino acids and contains several homologous regions in the alkaline phosphatases of E. coli, S.cerevisiae and human placenta.

• Journal of Microbiology and Biotechnology
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• 제13권6호
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• pp.846-852
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• 2003

rpoB DNAs (279 bp) from 34 species of 5 actinomycete genera were sequenced and a phylogenetic tree was constructed based on the sequences obtained. The genera were clearly differentiated in the rpoB tree, forming clades specific to their respective genus. In addition, 2 signature amino acid residues specific to Streptomyces were found in a multiple alignment of the deduced amino acid sequences. To empirically confirm that this rpoB gene analysis system could be used to differentiate actinomycete isolates, the proposed system was used to identify 16 actinomycete isolates from Jeju Island. All isolates were successfully differentiated into the genera Streptomyces and Micromonospora. Accordingly, this is the first report that an rpoB sequence analysis has been effectively used to differentiate actinomycete strains at the genus level.

• KSBB Journal
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• 제16권6호
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• pp.562-567
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• 2001

Styrene oxide 계열의 라세믹 에폭사이드 기질에 대한 입체선택적 가수분해능이 우수한 Aspergillus nigerr계열의 생촉매를 선발하였고, A.niger LK 유래의 EHase의 기질 특이성을 분석하였다. A. niger LK의 EHase는 benzene ring에 oxirane ring이 직접 연결되어 있는 styrene oxide, p-nitrostyrene oxide 기질에 대해서는 (R)-이성질체, benzene ring과 oxirane ring사이에 ether 등의 연결 chain이 있는 기질에 대해서는 (S)-이 성질체에 대한 입체선택적 가수분해능이 우수하였다. A niger LK의 EHase 유전자를 RT-PCR 방법으로 클로닝하였고, sequencing을 통해 다른 미생물 유래의 EHase와의 sequence identity 분석 등을 통해 특성을 분석하였다. Yeast 유래의 EHase와는 32% 수준의 sequence identity를 보였으며, Agrobacterisum, Corynebacterium 등의 박테리아 유래 EHase와는 identity가 매우 낮은 특성을 보였다. E. coli 숙주에서 발현된 재조합 EHase의 활성은 라세믹 에폭사이드 기질에 대한 입체선택적 가수분해 반응을 통해 확인할 수 있었다. 클러닝된 EHase의 보다 효율적인 발현 연구가 필요하며, 이러한 재조합 EHase는 고부가가치 광학활성 에폭사이드 제조를 위한 생물전환공정 시스템의 생촉매로 응용될 수 있을 것으로 기대된다.

• 식물조직배양학회지
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• 제26권1호
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• pp.41-47
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• 1999

토마토의 genomic DNA library로부터 분리한 tPALl, tPAL4유전자의 염기서열을 분석하여 tPAL5 유전자와 비교 분석한 결과는 다음과 같다. tPAL5 유전자는 722개의 아미노산과 710 bp의 intron을 가지고 있으나 tPALl은 intron을 가지고 있지 않으며 또한 tPAL5 유전자와 비교하여 249개의 짧은 polypeptide를 가지고 있었다. tPAL4유전자인 경우 357개의 아미노산과 305bp의 intron을 가지고 있었다. tPAL 효소간의 아미노산 homology는 tPAL1유전자와 tPAL4 유전자간은 87.2%, tPALl과 tPAL5는 85.3%, tPAL4 와 tPAL5 는 91.4%의 homology를 보였다. 또한, tPALl, tPAL4 유전자는 정상적인 polypeptide를 가지는 tPAL5유전자와 비교하여 비정상적인 stop codon을 가진 짧은 polypeptide로 구성되어 있었다. 다양한 식물 종으로부터 분리된 PAL유전자의 염기서열을 비교한 결과 토마토 (Lycopersicon esculentum), 감자 (Solanum tuberosum), 고구마 (Ipomoea batatas)간의 유연관계과 높았으며, parsley (Petroselinum crispum), bean (Phaseolus vulgaris), pea (Pisum sativum), alfalfa (Medicago sativa) 등이 각각 서로간에 유연관계가 높았다. 또한, 토마토에서 분리한 family내에서 tPAL4와 tPAL5 유전자는 homology가 매우 높았고 (93.0%), tPAL1와 tPAL4유전자 사이는 다소 낮았으며 (84.4%), 특히 tPAL4는 감자의 PAL 유전자와 매우 높았다 (90.6%).

• BMB Reports
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• 제31권6호
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• pp.542-547
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• 1998

The main function of adipocytes in various species is to store nutrient energy in the form of triglycerides, and this function may he closely related with hormonal signaling through the plasma membrane of adipocytes. Using SDS-PAGE, two-dimensional gel electrophoresis, and a membrane biotinylation technique, we have identified a 55KDa protein (55K protein) from the plasma membrane fraction of adipocytes, with an isoelectric point (pI) of 8.1-8.3. However, this 55K protein was not observed with a two-dimensional gel electrophoresis carried out on plasma membrane fractions prepared from the liver, heart, and kidney tissues. An analysis of the 12 amino acids sequence at the N-terminal of the 55K protein showed that it has a similar sequence to that of bovine serum albumin.

• Journal of Microbiology
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• 제39권2호
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• pp.109-115
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• 2001

Salmonella typhimurium possesses a cad operon, which contributes to an adaptive response against an acidifying environment. In Escherichia coli, the activation of the cad operon is dependent on cadC, which is located upstream of the operon. However, the activator of cad operon in S. typhimurium has not been known until now. In this study, we selected a putative cadC mutant by trasposon mutagenesis and cloned the cadc of S. typhimurium. Moreover, the cadC mutant was complemented by cadC clone. The cadC gene from S. typhimurium LT-2 consists of 1539 bp encoding a polypeptide ob 512 amino acids, and shows sequence similarity to cadC of E. coli with 53% identity and 67% similarity. The hydrophobicity profile of th S. typhimurim CadC sequence is very similar to E. coli CadC.

• Journal of Microbiology and Biotechnology
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• 제19권11호
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• pp.1306-1318
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• 2009

The filamentous ascomycete Sclerotinia sclerotiorum is well known for its ability to produce a large variety of hydrolytic enzymes. Two $\alpha$-amylases ScAmy54 and ScAmy43 predicted to play an important role in starch degradation were showed to produce specific oligosaccharides essentially maltotriose that have a considerable commercial interest. Primary structure of the two enzymes was established by N-terminal sequencing, MALDI-TOF masse spectrometry and cDNA cloning. The two proteins have the same N-terminal catalytic domain and ScAmy43 derived from ScAmy54 by truncation of 96 amino acids at the carboxyl-terminal region. Data of genomic analysis suggested that the two enzymes originated from the same $\alpha$-amylase gene and that truncation of ScAmy54 to ScAmy43 occurred probably during S. sclerotiorum cultivation. The structural gene of Scamy54 consisted of 9 exons and 8 introns, containing a single 1,500-bp open reading frame encoding 499 amino acids including a signal peptide of 21 residues. ScAmy54 exhibited high amino acid homology with other liquefying fungal $\alpha$-amylases essentially in the four conserved regions and in the putative catalytic triad. A 3D structure model of ScAmy54 and ScAmy43 was built using the 3-D structure of 2guy from A. niger as template. ScAmy54 is composed by three domains A, B, and C, including the well-known $(\beta/\alpha)_8$ barrel motif in domain A, have a typical structure of $\alpha$-amylase family, whereas ScAmy43 contained only tow domains A and B is the first fungal $\alpha$-amylase described until now with the smallest catalytic domain.