• Title/Summary/Keyword: Cell-of-origin

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Diagnosis of bovine cryptosporidiosis by indirect immunofluorescence assay using monoclonal antibody (단세포군항체를 이용한 간접형광항체법에 의한 송아지 작은와포자충증의 진단)

  • Wi, Seong-Hwan;Lee, Jeong-Gil;Ju, Hu-Don
    • Parasites, Hosts and Diseases
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    • v.33 no.2
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    • pp.107-116
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    • 1995
  • Two hybridoma cell lines against Cwptosporinium possum oocysts nFRl-CN911 were produced. The isotype of these 2 monoclonal antibodies (mAbs) was IgG2b (lE7.2) and and IgM (C6). Enzyme immuno-transfer blotting analysis showed that 157.2 reacted specifically to 36 kDa protein and C6 reacted to 67 and 70 kDa proteins. C. pcnlum was bound specifically to the surface region of oocysts by these mobs. No cross-reactivity was observed with tachyzoites of ToxopLosma gonnii and oocysts of Eimeria zuernii,5. bouis and E. canadensis of bovine origin. The indirect immunofluorescence assay (IIF) using mAb C6 was successful with counterstain. With the IIF using mob C6, oocysts appeared as 3 to $5{\mu}m$ spherical objects fluorescing bright apple green against a reddish dark background. The IIF using mAb C6 was agreed in specificity and sensitivity with those of a commercial diagnostic kit. These results demonstrated that the produced mAbs were specific to C. parvum and that the mAb C6 could be used for diagnosis of cryptosporidiosis.

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흑염소의 체내수정란 생산에 관한 연구

  • 최창용;조숙현;한만희;권응기;최성복;최연호;최순호;손동수;최상용
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.86-86
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    • 2002
  • 면양과 염소가 최근 수십년동안 세계여러 나라에서 번식생리의 연구를 위한 모델로 사용되어 왔는데, 체내수정란의 생산에 관한 영역도 유럽을 중심으로 활발하게 연구되어왔다. 수정란생산을 위한 발정동기화방법, 과배란처리 및 수정란회수방법 기술은 현재 상당히 많은 기술진척이 이루어진 상태이나, 우리나라 고유의 재래유전자원인 흑염소에는 이를 위한 기술이 미진한 실정이므로 본 실험에서는 흑염소의 체내수정란생산기술을 확립하여 재래가축 유전자원보존을 위한 기초기술을 마련하고자 한다. 축산기술연구소 남원지소에서 사육하고 있는 체중 20kg 이상의 건강한 흑염소를 이용하여 발정동기화를 위해 controlled intravaginal drug release(CIDR)를 질내에 14일 동안 삽입하고, 과배란처리는 FSH를 CIDR 삽입 12, 13, 14일째에 12시간 간격으로 점감법으로 총20mg을 투여하였으며, PGF$_2$a를 13일째 FSH와 함께 투여하였다. CIDR는 14일째의 아침에 제거하였다. 수컷과의 교미는 CIDR제거 24시간후에 GnRH를 투여와 동시에 실시하였으며, 채란은 교미후 3일째에 외과적인 방법으로 실시하였다. CIDR처리경과에 따른 progesterone농도는 CIDR 주입시 바로 수치가 상승하여 제거전까지 6~12ng/m1의 농도를 유지하였으며, 제거즉시 2ng/ml 이하로 떨어졌다. 채란시 평균 배란점은 16.5개, 미배란난포 9.8개였으며, 회수수정란은 6.0개를 나타내어 채란율은 36.4%를 나타내었다. 회수된 수정란의 발달단계는 4-cell 78.9%, 2-cell 5.3%, fragmentation 15.8%를 나타내었다. 이와 같은 체내수정란생산방법을 기반으로 하여 이후 수정란의 동결 및 수정란이식기법에 관한 연구를 수행한다면 우리나라의 재래가축인 흑염소의 유전자원 장기보존과 생산성향상에 기여할 것으로 사료된다.배양액에 30 embryos/50ul 소적으로하여 38.8$^{\circ}C$, 5% $CO_2$의 탄산가스 배양기에서 각각 7일간 배양을 실시하였다. 조사된 결과는 SAS/STAT를 이용하여 통계분석을 실시하였다. 체외수정 12시간 후에 난자 급속 염색법으로 염색을 실시한 결과, 모든 처리구에서 핵성숙률(76.4~95.2%), 정자침투율(51.1~66.9%), 웅성전핵형성률(95.2~100%), 다정자침입률(18.2~25.6%) 및 평균침입정자수(1.2~l.4개)에서 유의적인 차이가 인정되지 않았다. 체외배양 48시간 난할률을 조사한 결과, 처리구별 차이(53.9~67.9%)는 인정되지 않았으나, 배양 7일째 배반포형성률은 각각 14.5, 25.4, 17.3 및 12.4%로서 25uM의 $\beta$-ME처리구가 유의적(P<0.05)으로 높은 배발달률을 나타내었고, 총세포수에 있어서는 대조구와 처리구간 유의적인 차이가 인정되지 않았다. 따라서 돼지 난포란을 성숙배양할 때, 25uM $\beta$-ME를 첨가배양하는 것이 양질의 돼지체외수정란을 생산하는 하나의 방법으로 조사되었다.다.natural objects and was popular at the time of Yukjo Dynasty, and there are some documents of that period left both in Japan and Korea. "Hyojedo" in Korea is supposed to have been influenced by the letter design. Asite- is also considered to have been "Japanese Letter Jobcheso." Therefore, the purpose of this study is to look into the origin

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Mechanism of Anti-Invasive Action of Docosahexaenoic Acid in SW480 Human Colon Cancer Cell (인체 대장암 세포주 SW480에서 docosahexaenoic acid에 의한 침윤억제 기전)

  • Shin, So-Yeon;Kim, Yong-Jo;Song, Kyoung-Sub;Jing, Kaipeng;Kim, Na-Yeong;Jeong, So-Yeon;Park, Ji-Hoon;Seo, Kang-Sik;Heo, Jun-Young;Kwon, Hyun-Joo;Park, Jong-Il;Park, Seung-Kiel;Kweon, Gi-Ryang;Yoon, Wan-Hee;Hwang, Byung-Doo;Lim, Kyu
    • Journal of Life Science
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    • v.20 no.4
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    • pp.561-571
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    • 2010
  • Colon cancer is one of the most common malignancies in the western world and the second leading cause of cancer death in Korea. Epidemiology studies have shown a reduced incidence of colon cancer among populations consuming a large quantity of ${\omega}3$-polyunsaturated fatty acids (${\omega}3$-PUFA) of marine origin. Recently, it has been found that ${\omega}3$-PUFA has an antineoplastic effect in several cancers. This study was designed to investigate the mechanism of the anti-invasive effect of ${\omega}3$-PUFA in colon cancer. ${\omega}3$-PUFA, docosahexaenoic acids (DHA) and eicosapentaenoic acid (EPA) treatment resulted in a dose-dependent inhibition of cell growth in SW480 human colon cancer cells. In contrast, arachidonic acid (AA), a ${\omega}6$-PUFA, exhibited no significant effect. This action likely involves apoptosis, given that DHA treatment increased apoptotic cells in TUNEL assay. Moreover, invasiveness of SW480 cells was inhibited following treatment of DHA in a dose-dependent manner; in contrast, AA had no effect. The levels of MMP-9 and MMP-2 mRNA decreased after DHA pretreatment. MMP-9 and MMP-2 promoter activities were also inhibited by DHA treatment. The levels of NF-kB and p-IkB protein were down-regulated by DHA pretreatment in a dose dependent manner. In addition, DHA inhibited NF-kB promoter reporter activities. These findings suggest that ${\omega}3$-PUFA may inhibit cancer cell invasion by inhibition of MMPs via reduction of NF-kB in colon cancer. In conclusion, ${\omega}3$-PUFA could be used for chemoprevention and treatment of human colon cancer.

Mixed Osteosarcoma with Metastatic Alveolar Carcinomatous Appearance in Canine Mammary Gland Tumor (개의 유선에서 발생한 혼합성 골육종 및 샘포 상피세포의 전이성 암종)

  • Goo, Moon-Jung;Hong, Il-Hwa;Park, Jin-Kyu;Yang, Hai-Jie;Yuan, Dong-Wei;Ki, Mi-Ran;Lee, Hye-Rim;Hong, Kyung-Sook;Han, Jung-Youn;Hwang, Ok-Kyung;Kim, Tae-Hwan;Do, Sun-Hee;Jeong, Kyu-Shik
    • Journal of Life Science
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    • v.17 no.12
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    • pp.1766-1770
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    • 2007
  • We describe here a case of malignant mixed osteogenic tumor of the mammary gland with alveolar carcinomatous appreance. A firm, 2 to 2.5cm (in diameter) mass under the 5th nipple, showing the structure of extraosseous osteogenic sarcoma, was removed from the left 5th mammary gland of 12-year-old female dog. When investigated under the microscope, the osteoid material undergoing mineralization was surrounded by numerous scattered osteoblasts and a few osteoclastic cells throughout the osteoid tumorous stroma. The osteoid lesions were continuous with hypercellular myoepithelial cells of a very immature character with several mitotic figures. In addition, there were also carcinomatous tubules and alveoli, with invading cells into peripheral stroma, surrounded by myoepithelial cells in the mammary gland. In these lesions, emanating cords of tumor cells appear to be continuous with the myoepithelial cell layer of a duct. The presence of all these cell types suggests the existence of a common malignant origin, the stem cell being differentiated into epithelial carcinomatous and mesenchymal sarcomatous chondral and osteogenic tissues.

Quantification of endothelin-1 in human umbilical venous endothelial cell culture supernatants of small for gestational age and preeclampsia neonates (부당 경량아 및 전자간증 산모의 신생아 제대혈관내피세포의 endothelin-1 발현 비교)

  • Cho, Won Kyoung;Kim, So Young;Chun, Chung Sik
    • Clinical and Experimental Pediatrics
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    • v.50 no.12
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    • pp.1194-1199
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    • 2007
  • Purpose : It was generally accepted now a days that the pathogenesis of preeclampsia, small for gestational age (SGA), intrauterine growth retardation and fetal origin of adult diseases were related with a endothelial cell dysfunction. The purpose of this study was to know the relation of such diseases by assessing the level of endothelin-1. Methods : SGA babies, newborns of preeclampsia and normal control mother were included in this study. Isolated endothelial cells were centrifugated and mixed with media in $37^{\circ}C$, 5% $CO_2$ to obtain confluent monolayer of cultured human umbilical venous endothelial cell (HUVEC). Endothelin-1 levels were determined by Endothelin-1 colorimetric (EIA) Kits. We examined the endothelin-1 level in the HUVEC supernatants from SGA baby, and newborns from preeclampsia as well as normal mother. Also, we compared the endothelin-1 level of cultured normal HUVEC incubated with serum from cord blood of SGA, babies of preeclampsia or normal control mother. Results : The endothelin-1 levels in cultured HUVEC supernatants of three groups showed no significant difference but the endothelin-1 levels of cultured normal HUVEC incubated with serum from preeclampsia mother or SGA mother was significantly higher than those from newborns of control mothers (P<0.05). Conclusion : These findings suggest that there may be the factor which affect the endothelin-1 level in serum of cord blood from SGA and preeclampsia.

Soluble IL-2R, IFN-$\gamma$ and Neopterin as Immunologic Markers in Patients with Tuberculosis (결핵 환자에서 면역학적 지표로서의 sIL-2R, IFN-$\gamma$, Neopterin에 관한 연구)

  • Ryu, Yon-Ju;Ryu, Kum-Hei;Kim, Su-Hyun;Lee, Jong-Soo;Cheon, Seon-Hee;Seoh, Ju-Young
    • Tuberculosis and Respiratory Diseases
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    • v.53 no.3
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    • pp.294-308
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    • 2002
  • Background : The cell-mediated immune response plays an important role in tuberculosis. After being activated by mycobacterial antigens, T lymphocytes express a high affinity receptor (IL-2R) for interleukin-2 (IL-2) on their own surface and release a soluble fraction of the IL-2 receptor (sIL-2R) from the cell membrane into the circulation. Neopterin is a metabolite of guanosine-triphosphate, which is produced by stimulated macrophages under the influence of IFN-$\gamma$ with a T lymphocyte origin. Therefore, the utility of sIL-2R, IFN-$\gamma$ and the neopterin levels as immunologic indices of the cell-mediated immune response and severity of disease in patients with pulmonary tuberculosis was assessed. Methods : The serum sIL-2R, IFN-$\gamma$ and neopterin levels were measured in 39 patients with pulmonary tuberculosis, 6 patients with tuberculous lymphadenitis prior to treatment and 10 healthy subjects. The serum and pleural sIL-2R, neopterin and ADA levels were measured in 22 patients with tuberculous pleurisy. The patients with pulmonary tuberculosis were divided into a mild, moderate and severe group according to the severity by ATS guidelines. To compare the results from these patients with those of the pretreatment levels, the sIL-2R, IFN-$\gamma$ and neopterin levels were measured in 36 of the 39 patients(1 patient, expired; 2 patients were referred to a sanitarium) with pulmonary tuberculosis after 2 months of treatment. Results : 1) the serum sIL-2R and IFN-$\gamma$ levels were elevated in patients with tuberculosis when compared to those of healthy subjects (p>0.05). The neopterin concentration in the serum was significantly lower in patients with pulmonary tuberculosis($2967{\pm}2132.8$ pg/ml) than in healthy controls($4949{\pm}1242.1$ pg/ml)(p<0.05). 2) In the pulmonary tuberculosis group, the serum sIL-2R and IFN-$\gamma$ levels were higher in patients with severe disease than those in patients with mild and moderate disease. However, the neopterin levels declined as the pulmonary tuberculosis became more severe (p<0.01). 3) The mean serum sIL-2R and IFN-$\gamma$ levels declined from $1071{\pm}1139.4$ U/ml to $1023{\pm}1920.9$ U/ml(p>0.05), $41{\pm}52.8$ pg/ml to $22{\pm}23.9$ gm/ml(p<0.05), respectively, after 2 month of treatment. The mean serum neopterin levels increased from $3158{\pm}2272.6$ pg/ml to $3737{\pm}2307.5$ pg/ml(p>0.05) after a 2 month of treatment. These findings were remarkable in the severe group of pulmonary tuberculosis with a clinical correlation. 4) In the patients with tuberculous pleurisy, the serum sIL-2R and ADA were significantly higher than those in the pleural fluid, However, the neopterin levels in the sera and pleural effusion were similar. Conclusion : On the basis of this study, sIL-2R, IFN-$\gamma$ and neopterin measurements may not only provide an insight into the present state of the cell-mediated immune response, but also serve as parameters monitoring of the prognosis of the disease, particularly in patients with severe pulmonary tuberculosis. In addition, an assay of the pleural sIL-2R levels might signal a stimulated local immunity including T cell activation in the tuberculous pleural effusion.

EXPRESSION OF PROTEIN KINASE C ISOFORMS IN CHEMICAL CARCINOGEN-INDUCED NEOPLASTIC TRANSFORMATION OF HUMAN EPITHELIAL CELLS (화학적 발암화에 따른 Protein Kinase C의 발현 변화)

  • Byeon, Ki-Jeong;Hong, Lak-Won;Kim, Chin-Soo
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.23 no.4
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    • pp.295-305
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    • 2001
  • Protein kinase C (PKC) is known to play a pivotal role in neoplastic transformation cells and its high expression is often found in a variety of types of tumors including oral cancer. While PKC is associated with the altered signal transduction pathway of the tumor cells, it is still unclear which isoform is involved in the carcinogenesis process. Since the cellular distributions and the roles of PKC are isoform-specific, it is very important to identify the specific target molecules to improve our understanding of the carcinogenesis processes. Thus, the present study attempted to perform chemical carcinogen-induced neoplastic transformation of human epithelial cells and analyze the specific isoform of PKCs involved in the cellular transformation. The study analyzed overall PKC responses upon MNNG(N-Methyl-N'-nitro-N-nitroso guanidine) exposure with [$^3H$] PDBu binding assay. PKC translocation was observed at high doses of MNNG treatment in the presence of extracellular calcium. Such effects were not observed in the absence of extracellular calcium. Translocational effects with exposure of MNNG was further enhanced in the presence of hydrocortisone. The result suggests that the type of PKC involved may be $Ca^{2+}$-dependent classical isoform and steroid hormone enhances PKC activation. Among cPKC isoforms examined, only $PKC-{\alpha}$ and r showed significant translocation of protein levels from cytosolic fraction to membrane fraction, as analyzed by immunoblot. $PKC-{\varepsilon}$ in nPKC class showed an inch·eased translocation, but other forms in this class did not show the effect. None of isoforms in aPKC class was affected by MNNG treatment. The study demonstrated that there was a certain specificity in the patterns of isoform induction follwong chemical carcinogen exposure and helped identify all the types of PKC isoforms expressed in human epithelial cells. It was revealed that PKC isoforms were activated in an early resonse to chemical carcinogen, suggesting that PKC be associated with carcinogenesis process from an early stage in this particular cell system. The study will contribute to improving our understanding of chemical-induced carcinogenesis in human cells and may provide a scientific basis to introduce the specific PKC inhibitors as an anticancer drug of epithelial cell-origin cancers including oral cancer.

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Developmental Potentials of Clone Embryos Derived from Bovine Fetal Fibroblast Cells (소 태아섬유아세포 유래 복제란의 발육능에 관한 연구)

  • Cheong, H.T.;Kwon, D.J.;Park, Y.S.;Hwang, H.S.;Park, C.K.;Yang, B.K.;Kim, C.I.
    • Korean Journal of Animal Reproduction
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    • v.24 no.1
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    • pp.49-57
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    • 2000
  • This study was conducted to investigate the developmental potential of cloned embryos derived from bovine fetal fibroblast cells, and the effect of quiescent treatment, passage number and origin of donor cells on in vitro development of cloned embryos. Fetal skin and liver-derived fibroblast cells were transferred to enucleated oocytes after serum starvation or nontreatment (cycling). After electrofusion. reconstituted embryos were activated with $Ca^{++}$-ionophore and cycloheximide, and cocultured for 7~9 days with BRL cells. Some blastocysts were transferred to recipient cows 7~8 days post estrus. The development rate to the blastocyst stage of serum starved cell-derived embryos was higher (25.3%) than that of actively dividing cells-derived embryos (15.9%), The rates of blastocyst formation were 23.1~25.0% after transfer of cell passaged 4 to 6 times, and 23.8 and 25.2% after transfer of fetal skin and liver cells, respectively. After embryo transfer, 34.4% and 15.6% of recipient cows were pregnant on Day 60 and 120, respectively, and one male calf was produced from skin-derived vitrified blastocyst. The result of this study showed that the development of cloned embryos. was enhanced by quiescent treatment, but did not different among the cells passaged 4 to 6 times, and between skin and liver cells. This result also confirms that offspring can be obtained from the vitrified clone embryo derived from fetal skin cell.

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Evaluation of Commercial Korean Honey Quality and Correlation Analysis of the Quality Parameters (국내 시판 벌꿀의 품질 평가 및 품질인자간 상관관계 분석)

  • Sung, Hwa-Jung;Jung, Chuleui;Kwon, Jiyoung;Sohn, Ho-Yong
    • Journal of Life Science
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    • v.28 no.12
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    • pp.1489-1500
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    • 2018
  • Honey is made from flower nectar by honey bees. In this study, 120 honeys from various flowers and across eight different provinces in Korea were collected and their components, antioxidants, and hemolytic activities against red blood cell were evaluated. Our results show that total polyphenol (TP) varied widely across the samples, with chestnut honey showing the highest TP ($77.1{\pm}8.4mg/100g$), protein content ($25.9{\pm}0.9mg/100g$), and absorbance at 400 nm ($A_{400}$ : $0.156{\pm}0.036$). In contrast, the acacia honey and sugar honey had a TP of 9.5~30 mg, 12~15 mg/100g of, and the lowest $A_{400}$ of $0.06{\pm}0.02$. High amounts of total flavonoid were quantified in the jujube and chestnut honeys at $8.73{\pm}7.31$ and $8.39{\pm}3.02mg/100g$, respectively. No samples demonstrated hemolytic activity up to 1 mg/ml. Antioxidant activities determined by DPPH, ABTS, and nitrite scavenging placed the chestnut honey highest, followed by jujube, styrax, multi-floral, citrus, acacia and sugar honey. Analysis of parameter correlations indicated that the components and bioactivity of the honey are dependent on the origin of the flower rather than on bee-farming regions. A positive correlation between TP content and antioxidant activity was identified. The correlation coefficients between $A_{400}$ and the TP, ABTS scavenging, and reducing power values were 0.804, 0.772 and 0.741, respectively. We therefore suggest that $A_{400}$ could be used as a noble, economic and simple factor for honey quality evaluation. Our results can potentially be used to develop functional honey for the food and pharmaceutical industries.

Generation of antibodies against N-terminus fragment of AgI/II protein from Streptococcus mutans GS-5 (연쇄상구균(Streptococcus mutans GS-5)의 항원단백질 AgI/II의 N-terminus절편에 대한 항체형성)

  • Han, Ji-Hye;Baik, Byeong-Ju;Yang, Yeon-Mi;Park, Jeong-Yeol;Kim, Jae-Gon
    • Journal of the korean academy of Pediatric Dentistry
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    • v.33 no.3
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    • pp.401-410
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    • 2006
  • Dental caries results from localized demineralization of tooth enamel by acids of bacterial origin produced from the fermentation of dietary sugars. A group of related oral bacteria, collectively known as mutans streptococci, are implicated as the primary etiological agents of human caries. Within this group, Streptococcus mutans has been known as a causative agent for dental caries. As well as acid production yielding the demineralization of tooth enamel, adherence and colonization of S. mutans to the teeth are also important for their virulence Cell-surface fibrillar proteins, which mediate adherence to the salivary pellicle are virulence components of mutans streptococci, and primary candidates for a human caries vaccine. Here we report that the AgI/II gene from S. mutans GS-5 were cloned by PCR amplification of the bacterial chromosomal DNA and the integrity of cloned genes were confirmed by nucleotide sequencing. Sequence analyses showed the sequence alignment of 280 nucleotides between the cloned AgI/II and the reported sequence of S. mutans GS-5 showed the perfect match The cloned genes which signal nucleotide was truncated, were transferred into bacterial expression vector and the recombinant proteins were purified as His-tag fusion proteins In order to generate polyclonal antibodies against the recombinant proteins, AgI/II mr, some $100{\mu}g$ of the proteins was injected into mice three times. It can be used for an effective vaccine production to prevent dental caries caused by pathogenic S. mutans.

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