• Parasites, Hosts and Diseases
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• 제49권1호
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• pp.79-83
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• 2011

Trichomoniasis is a sexually transmitted disease due to infection with Trichomonas vaginalis, and it can cause serious consequences for women's health. To study the virulence factors of this pathogen, T. vaginalis surface proteins were investigated using polyclonal antibodies specific to the membrane fractions of T. vaginalis. The T. vaginalis expression library was constructed by cloning the cDNA derived from mRNA of T. vaginalis into a phage ${\lambda}$ Uni-ZAP XR vector, and then used for immunoscreening with the anti-membrane proteins of T. vaginalis antibodies. The immunoreactive proteins identified included adhesion protein AP65-1, ${\alpha$-actinin, kinesin-associated protein, teneurin, and 2 independent hypothetical proteins. Immunofluorescence assays showed that AP65-1, one of the identified immunogenic clones, is prevalent in the whole body of T. vaginalis. This study led us to identify T. vaginalis proteins which may stimulate immune responses by human cells.

• 대한수의학회지
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• 제39권4호
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• pp.797-805
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• 1999

T sergenti cDNA library were constructed to get a more broad information about the structural, functional or antigenic properties of the proteins, and analyzes for their partial cDNA sequences and expression sequences tags(ESTg). The mRNA were purified from T sergenti isolates to identify the information of antigen gene, then first and second strand cDNA was synthesized. EcoR I adaptor ligation and Xho I enzyme restriction were used to the synthesized cDNA, and ligated into a Uni-ZAP XR vector. T sergenti cDNA library was constructed with packaging and amplification in vitro. Antibody screening was performed with constructed T sergenti cDNA library using antisera against T sergenti. Among those clones, eight phagemids were rescued from the recombinant in vivo excision with f1 helper phage. Using the analysis of endonuclease restriction and PCR, the recombinant cDNA were proved having a 0.5-3.0kb of inserts. The eight of partial cDNA clones' sequences were obtained and examined for their homology using BLASTN and BLASTX. The eight of sequenced clones were classified into three groups according to the basis of database searches. A total 3,045bp of partial cDNA sequence were determined from six clones. The putatively identified clones contain a cytochrome c gene, a heat shock protein gene, a cyclophilin gene, and a ribosomal protein gene. The unidentified clones have a homology to ATP-binding protein(mtrA) gene of S argillaceus, DNA-binding protein(DBP) gene of Pseudorabies virus 85kDa merozoite protein gene of B bovis, mRNA spm1 protein of T annulata and glycine-rich RNA-binding protein mRNA of O sativa etc.

• 한국잠사곤충학회지
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• 제38권1호
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• pp.19-24
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• 1996

누에에서 생체 방어에 관련된 새로운 항 세균성 펩타이드 유전자를 탐색 분리하기 위하여 누에 체강에 비 병원성 세균인 Escherichia coli를 주사하여 면역 반응의 일환으로 발현량이 증가하는 유도 유전자 종류를 조사 하였다. 체강 주사 8시간 후 누에에서 cDNA 유전자 은행을 만들고, 정상 및 유도 누에에서 분리한 각각의 mRNA를 탐침으로 차별화 선별을 하였다. 차별화 선별 결과 정상보다도 유도 누에의 탐침을 사용한 막에서 강도가 높은 클론 32개를 선발하였고, 29개 클론에 대해 전체 또는 부분 염기 서열을 분석하여 DNA 상동성을 조사하였다. DNA 상동성 비교를 통해 생산한 발현 유전자 꼬리표 중에는 비교적 상동 유의성이 인정되어 그 실체를 추정할 수 있는 19개의 클론이 있었다. 특히 곤충의 면역 작용에 직접적으로 관계하는 항세균성 펩타이드 유전자, hemolin 유전자, transferrin 유전자 등 4종의 유전 자원을 확보할 수 있었다.

• Journal of Applied Biological Chemistry
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• 제44권2호
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• pp.59-64
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• 2001

Pepper plants (Nogkwang, 60-day old) were inoculated with Phytophthora capsici to induce sesquiterpene cyclase associated with the biosynthesis of phytoalexin (capsidiol), a substance related to the defense against pathogens in plants. One day after inoculation, mRNA was isolated from the root, cDNA synthesized, and a library constructed in a ZAP express XR vector. The efficiency was $2{\times}10^6pfu/{\mu}g$. Sesquiterpene cyclase cDNA from Hyoscyamus muticus was labeled with $^{32}P$ and used as a probe for screening the cDNA library. After the third screening, 25 positive clones were selected. Through restrictive digestion and DNA gel-blot analysis, six different cyclase gene expressions were identified. PSC1B sequences of the six clones were determined, which were 1966 base pairs encoded 556 amino acids with an expected molecular weight of 63.8 kDa. Response against the pathogen was different between the resistant and susceptible peppers. After the infection of the pathogen, the expression of PSC genes continued in the resistant peppers while the plants were alive. The expression in the susceptible peppers lasted for only 4 days.

• Genomics & Informatics
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• 제21권2호
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• pp.18.1-18.11
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• 2023

Immunologists have activated T cells in vitro using various stimulation methods, including phorbol myristate acetate (PMA)/ionomycin and αCD3/αCD28 agonistic antibodies. PMA stimulates protein kinase C, activating nuclear factor-κB, and ionomycin increases intracellular calcium levels, resulting in activation of nuclear factor of activated T cell. In contrast, αCD3/αCD28 agonistic antibodies activate T cells through ZAP-70, which phosphorylates linker for activation of T cell and SH2-domain-containing leukocyte protein of 76 kD. However, despite the use of these two different in vitro T cell activation methods for decades, the differential effects of chemical-based and antibody-based activation of primary human T cells have not yet been comprehensively described. Using single-cell RNA sequencing (scRNA-seq) technologies to analyze gene expression unbiasedly at the single-cell level, we compared the transcriptomic profiles of the non-physiological and physiological activation methods on human peripheral blood mononuclear cell-derived T cells from four independent donors. Remarkable transcriptomic differences in the expression of cytokines and their respective receptors were identified. We also identified activated CD4 T cell subsets (CD55⁺) enriched specifically by PMA/ionomycin activation. We believe this activated human T cell transcriptome atlas derived from two different activation methods will enhance our understanding, highlight the optimal use of these two in vitro T cell activation assays, and be applied as a reference standard when analyzing activated specific disease-originated T cells through scRNA-seq.

• International Journal of Industrial Entomology and Biomaterials
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• 제28권2호
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• pp.71-84
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• 2014

The full-length heat shock protein 88 (HSP88) complementary DNA (cDNA) of Paecilomyces tenuipes Jocheon-1 was obtained by screening the Paecilomyces tenuipes (P. tenuipes) Jocheon-1 Uni-Zap cDNA library and performing 5' RACE polymerase chain reaction (PCR). The P. tenuipes Jocheon-1 HSP88 cDNA contained an open reading frame (ORF) of 2,139-basepair encoding 713 amino acid residues. The deduced amino acid sequence of the P. tenuipe s Jocheon-1 HSP88 cDNA showed 77% identity to Nectria haematococca HSP88 and 45-76% identity to other fungal homologous HSP88s. Phylogenetic analysis and BLAST program analysis confirmed that the deduced amino acid sequences of the P. tenuipes Jocheon-1 HSP88 gene belonged to the ascomycetes group within the fungal clade. The P. tenuipes Jocheon-1 HSP88 also contained the conserved ATPase domain at the N-terminal region. The cDNA encoding P. tenuipes Jocheon-1 HSP88 was expressed as an 88 kilodalton (kDa) polypeptide in baculovirus-infected insect Sf9 cells. Under higher temperature conditions for the growth of the entomopathogenic fungus, mRNA expression of P. tenuipes Jocheon-1 HSP88 was quantified by real time PCR (qPCR). The results showed that heat shock stress induced a higher level of mRNA expression compared to normal growth conditions.

• Journal of Microbiology
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• 제44권1호
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• pp.54-63
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• 2006

Ferritin is a major eukaryotic protein and in humans is the protein of iron storage. A partial gene fragment of ferritin (255 bp) taken from the total RNA of Periserrula leucophryna, was amplified by RT-PCR using oligonucleotide primers designed from the conserved metal binding domain of eukaryotic ferritin and confirmed by DNA sequencing. Using the $^{32}P-labeled$ partial ferritin cDNA fragment, 28 different clones were obtained by the screening of the P. leucophryna cDNA library prepared in the Uni-ZAP XR vector, sequenced and characterized. The longest clone was named the PLF (Periserrula leucophryna ferritin) gene and the nucleotide and amino acid sequences of this novel gene were deposited in the GenBank databases with accession numbers DQ207752 and ABA55730, respectively. The entire cDNA of PLF clone was 1109 bp (CDS: 129-653), including a coding nucleotide sequence of 525 bp, a 5' -untranslated region of 128 bp, and a 3'-noncoding region of 456 bp. The 5'-UTR contains a putative iron responsive element (IRE) sequence. Ferritin has an open reading frame encoding a polypeptide of 174 amino acids including a hydrophobic signal peptide of 17 amino acids. The predicted molecular weights of the immature and mature ferritin were calculated to be 20.3 kDa and 18.2 kDa, respectively. The region encoding the mature ferritin was subcloned into the pT7-7 expression vector after PCR amplification using the designed primers and included the initiation and termination codons; the recombinant clones were expressed in E. coli BL21(DE3) or E. coli BL21(DE3)pLysE. SDS-PAGE and western blot analysis showed that a ferritin of approximately 18 kDa (mature form) was produced and that by iron staining in native PAGE, it is likely that the recombinant ferritin is correctly folded and assembled into a homopolymer composed of a single subunit.

• Clinical and Experimental Reproductive Medicine
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• 제25권2호
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• pp.123-128
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• 1998

Implantation rates remain low following human in vitro fertilization (IVF). Suboptimal culture conditions may limit the ability of embryos to hatch as blastocysts, and artificial opening of the zona pellucida has been proposed as a means to promote subsequent hatching (assisted hatching). In this study, assisted hatching (AH) by zona drilling using acidic Tyrode's solution was performed in 320 patients, due to their age of more than 38 years (group A), the thick zona pellucida (group Z; $ZP\geq0.18{\mu}m$), and failures in implantation more than 3 times in previous IVF-ET trial (group P). This study was designed firstly, to study the effects of AH on the outcomes of IVF-ET according to the indications and secondly, to verify the appropriate application of AH. The results were as follows; 1. There was no difference in pregnancy rate between AH group (26.6%) and non-AH group (26.5%). 2. Assisted hatching (AH) showed significantly higher pregnancy rate of the patients with thick zona pellucid a than those of the patients with age factor and with the history of repeated implantation failure. But in the patients with age factor only, AH resulted in higher pregnancy rate. 3. Interestingly, the patients with complex factors including zona factor (Z: 33.9%; ZA: 30.4%; ZP: 31.6%; ZAP: 21.4%) showed higher pregnancy rates than other complex factors excluding zona factor (A: 24.4%, P: 0%; AP: 10.8%). From these results, AH is more helpful to the patients with thick zona pellucida rather than patients with older age and/or previous repeated implantation failure.

• 디자인학연구
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• 제18권1호
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• pp.39-48
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• 2005

지금의 디지털 문화에서 TV 방송은 정보전달과 커뮤니케이션 매체로서 과거보다 더 큰 영향력을 발휘하게 되었다. 우리나라 방송환경은 2002년 위성방송의 출범으로 지상파, 케이블, 위성, 인터넷 등으로 다원화됨으로써 다매체, 다채널 시대로 진입되었다. 이 같은 방송 환경의 변화는 TV 시청자들을 과거 수동적 이미지의 수용자에서 능동적인 커뮤니케이션의 주체자, 참여자, 이용자로서 미디어를 선택하고 이용하는 존재라는 새로운 수용자로 출현시켰다. 본 연구는 채널선택의 주체로 능동적 수용자가 방송의 중심에 서게 되면서 방송 프로그램의 자기 정체성에 대한 문제제기를 브랜드를 통해 구조화시키고자 한다. 능동적 수용자에 대한 인지심리학적 접근방법 가운데 근래 크게 주목받는 이론이 '스키마(uh,:ma)이론'이다. 이야기 스키마를 적용하여 '정보처리과정(information processing)'과 연관되어 있는 '인지적 과정(cognitive processes)'을 설명하는데 있어 브랜드의 활용은 TV 방송 프로그램이라는 원형의 각 부분을 추상으로 나타낼 수 있다. 이것은 수용자들이 선호 프로그램을 기억하고 재핑(Zapping)하는 시청습관의 형성 위에서 수용자 만족과 프로그램의 인지도 향상을 위한 중요한 변인을 브랜드로 인식하고 TV방송 프로그램의 전문 브랜드를 개발하는 실증연구로 이어진다. 연구범위는 TV 뉴스 프로그램을 대상으로 2004년 3월에서 5월에 걸쳐 방송중인 뉴스 프로그램을 국내외 각 방송사별로 조사하였으며, 보편적인 뉴스 전문 프로그램을 중심으로 진행하였다. 본 연구를 통해 TV 뉴스 프로그램의 경우, 방송사를 대표할 수 있는 전문 브랜드의 개발과 함께 뉴스앵커의 브랜드화 역시 중요한 변인이 될 수 있음을 확인할 수 있었다.지에 대한 고유한 특성을 파악할 수 있었다.인 기여가 큰 것으로 평가되었다. 퇴적물은 공간적으로 다소 불규칙한 변동이 있지만, 저질조성과 관련해서 양자강 하구쪽으로 갈수록 오염도가 낮고, 조사해역의 동쪽부분에서 유기오염의 정도가 상대적으로 크게 나타났지만, 그 오염도는 심하지 않는 것으로 분석되었다. 향후 본 대상해역에 있어서 산샤댐 건설과 관련된 양자강 유량의 변화, 그리고 어업활동과 관련된 폐기물에 의한 해양환경의 영향평가 및 대책 연구가 수행되어져야 할 것이다. 시도는 긍정적으로 평가되어야 한다. 그러므로 앞으로는 제품디자인 뿐만이 아닌 다른 다양한 분야들에게로 그 범위가 확대되어 디자인 문화적 정체성을 확립하는 연구가 뒤따라야 할 것이다.. 즉, 제품디자인의 결정요인 분석결과는 QFD의 접근방법에, 제품 디자인 파급효과 분석결과는 컨조인트 분석에 각각 보완적 기여를 할 수 있다. 이와 동시에, 실증적 분석결과는 Ettlie(1997)의 디자인 통합(DI) 이론에 대한 실증적 기반을 제공할 수 있다. 마지막으로, 성공적인 디자인 경영(DM)을 위해서는 최고 경영자의 지원뿐만 아니라 부처 간 의사소통의 장애요인을 제거하고 CFT(cross-functional team)를 운영함으로써 동시적 엔지니어링(CE) 및 제품 및 공정 디자인의 개발이 제품 개발의 속도를 가속화하고 디자인 품질을 높이며 시장 성공을 보증할 수 있도록 해야 한다.임과 채팅은 긍정적인 상호관련을 가진 것으로 나타난 반면 전자메일 서비스 이용은 성적 만족과 부정적인 상호관련을 가진 것으로 분석되었다. 이는 대학생들이 지루하게 느끼거나 외로움을 느낄 때 전자메일을 주로 이용하지만 성적 만족을 위해 전자메일을 이용하지 않고 있다는 사실을 보여주는 것이다. (3) 인터넷 이용