• Title/Summary/Keyword: Sequence to Sequence

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ANALYSIS OF DIFFERENTIAL GENE EXPRESSION IN NORMAL, CYST AND AMELOBLASTOMA CELLS (정상, 낭종 및 법랑아세포종 세포에서의 유전자 발현 차이 분석)

  • Yang, Cheol-Hee;Baik, Byeong-Ju;Yang, Yeon-Mi;Kim, Jae-Gon
    • Journal of the korean academy of Pediatric Dentistry
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    • v.32 no.1
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    • pp.75-88
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    • 2005
  • Ameloblastoma is the most commonly occurring odontogenic tumor in oral cavity. Although most are benign epithelial neoplasm, they are generally considered to be locally aggressive and destructive, exhibiting a high rate of recurrence. The biological behavior of this neoplasm is a slowly growing, locally invasive tumor without metastasis, therefore malignant neoplasm, changed its histological appearance to carcinoma or showed distant metastasis, is only defined clinically. In this study, we identified the differentially expressed genes(DEGs) in stages under benign or malignant ameloblastoma compared with normal patient using ordered differential display(ODD) reverse transcription(RT)-PCR and $GeneFishing^{TM}$ technology. ODD RT-PCR is rather effective when the investigation of samples containing very small amounts of total RNA must be accomplished. ODD RT-PCR used the means of amplification with anchored T-primer and adaptor specific primer. bearing definite two bases at their 3' ends and so this method could display differential 3'-expressed sequence taqs(ESTs) patterns without using full-length cDNAs. Compared with standard differential display, ODD RT-PCR is more simple and have enough sensitivity to search for molecular markers by comparing gene expression profiles, However, this method required much effort and skill to perform. $GeneFishing^{TM}$ modified from DD-PCR is an improved method for detecting differentially expressed genes in two or more related samples. This two step RT-PCR method uses a constant reverse primer(anchor ACP-T) to prime the RT reaction and arbitrary primer pairs(annealing control primers, ACPs) during PCR. Because of high annealing specificity of ACPs than ODD RT-PCR, the application of $GeneFishing^{TM}$ to DEG discovery generates reproducible, authentic, and long(100bp to 2kb) PCR products that are detectable on agarose gels. Consequently, various DEGs observed differential expression levels on agarose gels were isolated from normal, benign, and malignant tissues using these methods. The expression patterns of the some isolated DEGs through ODD RT-PCR and $GeneFishing^{TM}$ were confirmed by Northern blot analysis and RT-PCR. The results showed that these identified DEGs were implicated in ameloblastoma neoplasm processes. Therefore, the identified DEGs will be further studied in order to be applied in candidate selection for marker as an early diagnosis during ameloblastoma neoplasm processes.

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Characters of motile aeromonads isolated from imported ornamental fish (수입 관상어에서 분리한 motile aeromonads의 특성)

  • Jin, Se-yun;Ko, Chang-yong;Lee, Ye-ji;Jung, Yun-hee;Ju, Seong-cheol;Kim, Eunheui
    • Journal of fish pathology
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    • v.30 no.2
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    • pp.89-96
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    • 2017
  • The majority of freshwater ornamental fish are imported and distributed domestically, causing high risk of exposure to exotic pathogens and drug resistant bacteria in Korea. Aeromonas hydrophila is known as a common species of fresh water bacteria and opportunistic fish pathogen, as well as a species causing zoonotic infection. In this study, we isolated motile aeromonads from various imported freshwater ornamental fish and studied the characters of the isolates. Imported freshwater ornamental fish were purchased on day 1 after the fish were deposited in the aquarium. Bacteria were isolated from the liver, kidney and spleen of fish using 0.5% NaCl containing tryptic soy agar medium. Bacteria were grouped on the basis of their morphological characteristics. The colonies with clear zone on starch-ampicillin agar (SA agar) were tentatively identified as Aeromonas spp. Two hundred and twenty-six strains, about 70% of total isolates were assumed to be Aeromonas spp. Nine isolates were further identified based on the result of the API 20E test and PCR using primers specific for A. hydrophila 16S rRNA gene. The isolates were identified as A. hydrophila and the API 20E test showed differences in trisodium citrate, D-sucrose, D-melibiose, amygdalin and L-arabinose availability between the nine isolates and standard A. hydrophila. The susceptibilities of the isolated bacteria to 10 antibacterial agents were confirmed by the disk diffusion method. Isolated strains were found to be resistant to amoxicillin and ampicillin and sensitive to florfenicol. However, 7 isolates showed multiple drug resistances to erythromycin, oxytetracycline, nalidixic acid etc. Pathogenicity of the isolates was determined by the artificial challenge test on goldfish (Carassius auratus). Three isolates caused 60 ~ 80% mortality in goldfish within 5 days after the initiation of challenge. These results indicate that multiple drug resistant, highly pathogenic and exotic A. hydrophila can spread to domestic aquarium and the preventive treatment of fish before sale is necessary.

THE EFFECT OF ALTERED FUNCTIONAL FORCE ON THE EXPRESSION OF SPECIFIC MRNAS IN THE DEVELOPING MOUSE MANDIBLE (하악골의 발육중인 생쥐에서 기능력의 변화가 특이-유전자 발현에 미치는 영향)

  • Kim, Hyung-Tae;Park, Joo-Cheol;Lee, Chang-Seop;Park, Heon-Dong
    • Journal of the korean academy of Pediatric Dentistry
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    • v.30 no.2
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    • pp.308-319
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    • 2003
  • Mechanical forces are known to have an effect on bone formation, maintenance and remodeling, and there is evidence that the development of the mandibular condyle in the rat or mouse is influenced by altered functional force. However, studies are lacking in molecular-biologic mechanism such as the identification of differentiation factor induced from functional force. Here a mouse model was used to investigate the functional stress-responsive gene or factors which is related to the altered force by comparing the expression genes of functional state and hypo-functional state of the mouse mandible. ICR mice were provisioned with either a soft, mushy diet (soft-diet group) or hard rat pellets (hard-diet group) beginning at weaning for the alteration of functional force and subsequently sacrificed at 89 days of age. Incisor of mice in group 1 were trimmed twice a week to reduce occlusal forces. After killing the animals, mandibular bone including condyle were collected for RNA extraction, subtractive hybridization, northern blot analysis and mRNA in-situ hybridization. The results as follows; 1. A total of 39 clones were sequenced, and 11 individual sequence types were subsequently identified by subtractive hybridization, as 28 clones were represented twice in the analyzed sets. 2. Consequently four candidate clones, FS-s (functional stress-specific)2, -5, -18, and -22 were identified and characterized by homolgy search and northern analysis. Four of these clones, FS-s2, -5, -18, and -22, were shown to be expressed differentially in the hard-diet group. 3. Histologic sections showed that osteoblastic activity along the bone trabeculae and active bone remodeling were significantly lower in soft than in hard diet animals. A soft diet seems to enable a longer period of endochondral ossification in the mandibular condyle. 4. Although the mRNAs of FS-s2, -5, -18, and -22 were expressed rarely by cells of the soft-diet group, highest expression was detected in the cells of the hard-diet group. Together with the above results, it is suggested that FS-s2, -5, -18, and -22 could act as an important factors controlling the tissue changes in response to functional stress. The exact functional significance of these findings remains to be established.

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Voxel-based Investigations of Phase Mask Effects on Susceptibility Weighted Images (화소 간 분석을 이용하여 자화율 가중 영상(SWI)에 나타난 위상 마스킹의 효과 분석)

  • Hwang, Eo-Jin;Kim, Min-Ji;Kim, Hyug-Gi;Ryu, Chang-Woo;Jahng, Geon-Ho
    • Progress in Medical Physics
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    • v.24 no.1
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    • pp.25-34
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    • 2013
  • To investigate effects of phase mask on susceptibility-weighted images (SWI) using voxel-based analyses in normal elderly subjects. A three-dimensional (3D) gradient echo sequence ran to obtain SWIs in 20 healthy elderly subjects. SWIs with two (SWI2) and four (SWI4) phase multiplications were achieved with positive (PSWI) and negative (NSWI) phase masks to investigate phase mask effects. The voxel-based comparisons were performed using paired t-tests between PSWI and NSWI and between SWI2 and SWI4. Differences of signal intensities between magnitude images and SWI4 were larger than those between magnitude images and SWI2s. Differences of signal intensities between magnitude images and PSWIs were larger than those between magnitude images and NSWIs. Moreover, the signal intensities from NSWI2s and NSWI4s were greater than those from PSWI2s and PSWI4s, respectively. More differences of signal intensities between NSWI4 and PSWI4s were found than those between NSWI2s and PSWI2s in the whole brain images. The voxel-based analyses of SWI could be beneficial to investigate susceptibility differences on the entire brain areas. The phase masking method could be chosen to enhance brain tissue contrast rather than to enhance venous blood vessels. Therefore, it is recommended to apply voxel-based analyses of SWI to investigate clinical applications.

Distribution and Stratigraphical Significance of the Haengmae Formation in Pyeongchang and Jeongseon areas, South Korea (평창-정선 일대 "행매층"의 분포와 층서적 의의)

  • Kim, Namsoo;Choi, Sung-Ja;Song, Yungoo;Park, Chaewon;Chwae, Ueechan;Yi, Keewook
    • Economic and Environmental Geology
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    • v.53 no.4
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    • pp.383-395
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    • 2020
  • The stratigraphical position of the Haengmae Formation can provide clues towards solving the hot issue on the Silurian formation, also known as Hoedongri Formation. Since the 2010s, there have been several reports denying the Haengmae Formation as a lithostratigraphic unit. This study aimed to clarify the lithostratigraphic and chronostratigraphic significance of the Haengmae Formation. The distribution and structural geometry of the Haengmae Formation were studied through geologic mapping, and the correlation of relative geologic age and the absolute age was performed through conodont biostratigraphy and zircon U-Pb dating respectively. The representative rock of the Haengmae Formation is massive and yellow-yellowish brown pebble-bearing carbonate rocks with a granular texture similar to sandstone. Its surface is rough with a considerable amount of pores. By studying the mineral composition, contents, and microstructure of the rocks, they have been classified as pebble-bearing clastic rocks composed of dolomite pebbles and matrix. They chiefly comprise of euhedral or subhedral dolomite, and rounded, well-sorted fine-grained quartz, which are continuously distributed in the study area from Biryong-dong to Pyeongan-ri. Bedding attitude and the thickness of the Haengmae Formation are similar to that of the Hoedongri Formation in the north-eastern area (Biryong-dong to Haengmae-dong). The dip-direction attitudes were maintained 340°/15° from Biryong-dong to Haengmae-dong with a thickness of ca. 200 m. However, around the southwest of the studied area, the attitude is suddenly changed and the stratigraphic sequence is in disorder because of fold and thrust. Consequently, the formation is exposed to a wide low-relief area of 1.5 km × 2.5 km. Zircon U-Pb age dating results ranged from 470 to 449 Ma, which indicates that the Haengmae Formation formed during the Upper Ordovician or later. The pebble-bearing carbonate rock consisted of clastic sediments, suggesting that the Middle Ordovician conodonts from the Haengmae Formation must be reworked. Therefore, the above-stated evidence supports that the geologic age of the Haengmae Formation should be Upper Ordovician or later. This study revealed that the Haengmae Formation is neither shear zone, nor an upper part of the Jeongseon Limestone, and is also not the same age as the Jeongseon Limestone. Furthermore, it was confirmed that the Haengmae Formation should be considered a unit of lithostratigraphy in accordance with the stratigraphic guide of the International Commission on Stratigraphy (ICS).

Ore Mineralization of The Copper-bearing Hanae Hydrothermal Vein Deposit (하내 함 동 열수 맥상광상의 광화작용)

  • Choi, Sang-Hoon;Lee, Sunjin;Jun, Youngshik
    • Economic and Environmental Geology
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    • v.50 no.6
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    • pp.435-443
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    • 2017
  • The Hanae deposit is located within the Cretaceous Gyeongsang Basin. The Cu-bearing hydrothermal quartz vein formed by narrow open-space filling along fracture in the sedimentary rocks as Jindong Formation. The Hanae Cu-bearing hydrothermal deposit shows a paragenetic sequence of pyrrhotite-pyrite $\rightarrow$ pyrite-chalcopyrite-sphalerite(${\pm}$Bi-bearing tellurides) $\rightarrow$ Ag-bearing telluride mineralization $\rightarrow$ secondary mineralization. Fluid inclusion data indicate that the Hanae Cu-bearing hydrothermal mineralization occurred from dominantly aqueous fluids at temperatures of $400^{\circ}C-200^{\circ}C$. Equilibrium thermodynamic interpretation of the mineral paragenesis and assemblages combined with fluid inclusion data indicate that early main Cu-bearing ore mineralization in the vein starts at about $350^{\circ}C$ which corresponds to sulfur fugacity from about $10^{-9.2}$ to $10^{-8.7}bar$ with oxygen fugacity of about $10^{-32.1}$ to $10^{-29.8}bar$. Late main Cu-bearing ore mineralization in the vein occurs at about $250^{\circ}C$ which corresponds to sulfur fugacity from about $10^{-13.5}$ to $10^{-11.7}bar$ with oxygen fugacity of about $10^{-38.4}$ to $10^{-35.2}bar$. The late Ag-bearing telluride mineralization in the Hanae hydrothermal system occurs at about $200^{\circ}C$ which corresponds to minium Tellirium fugacity value of about $10^{-18}bar$ with sulfur fugacity of about $10^{-14.0}$ to $10^{-10.9}bar$.

Molecular Characterization and Phylogenetic Analysis of Season Influenza Virus Isolated in Busan during the 2006-2008 Seasons (부산지역에서 유행한 계절인플루엔자바이러스의 유전자 특성 및 계통분석('06-'08 절기))

  • Park, Yon-Koung;Kim, Nam-Ho;Choi, Seung-Hwa;Lee, Mi-Oak;Min, Sang-Kee;Kim, Seong-Joon;Cho, Kyung-Soon;Na, Young-Nan
    • Journal of Life Science
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    • v.20 no.3
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    • pp.365-373
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    • 2010
  • To monitor newly emerged influenza virus variants and to investigate the prevalence pattern, our laboratory performed isolation of the viruses from surveillance sentinel hospitals. In the present study, we analysed influenza A/H1N1, A/H3N2, B viruses isolated in Busan during the 2006/07 and 2007/08 seasons by sequence analysis of the hemagglutinin (HA1 subunit) and neuraminidase (NA) genes. The isolates studied here were selected by the stratified random sample method from a total of 277 isolates, in which 15 were A/H1N1, 16 were A/H3N2 and 29 were B. Based on the phylogenetic tree, the HA1 gene showed that A/H1N1 isolates had a 96.7% to 97.7% homology with the A/Brisbane/59/2007, A/H3N2 isolates had a 98.4% to 99.7% homology with the A/Brisbane/10/2007, and B isolates had a 96.5% to 99.7% homology with the B/Florida/4/2006(Yamagata lineage), which are all the vaccine strains for the Northern Hemisphere in 2008~2009 season. In the case of the NA gene, A/H1N1 isolates had 97.8% to 98.5% homologies, A/H3N2 isolates had 98.9% to 99.4% homologies, and B isolates had 98.9% to 100% homologies with each vaccine strain in the 2008~2009 season, respectively. Characterization of the hemagglutinin gene revealed that amino acids at the receptor-binding site and N-linked glycosylation site were highly conserved. These results provide useful information for the control of influenza viruses in Busan and for a better understanding of vaccine strain selection.

Clinical Utility of Turbo Contrase-Enhanced MR Angiography for the Major Branches of the Aortic Arch (대동맥궁 주요 분지들의 고속 조영증강 자기공명혈관조영술의 임상적 유용성)

  • Su Ok Seong
    • Investigative Magnetic Resonance Imaging
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    • v.2 no.1
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    • pp.96-103
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    • 1998
  • Purpose : To assess the clinical utility of turbo contrast-enhanced magnetic resonance angiography(CE MRA) in the evaluation of the aortic arch and its major branches and to compare the image quality of CE MRA among different coils used. Materials and Methods : Turbo three-phase dynamic CE MRA encompassing aortic arch and its major branches was prospectively performed after manual bolus IV injection of contrast material in 29 patients with suspected cerebrovascular diseases at 1.0T MR unit. the raw data were obtained with 3-D FISH sequence (TR 5.4ms, TE 2.3ms, flip angle 30, slab thickness 80nm, effective slice thickness 4.0mm, matrix size $100{\times}256$, FOV 280mm). Total data acquisition time was 4. to 60 seconds. We subjectively evaluated the imge quality with three-rating scheme : "good" for unequivocal normal finding, "fair" for relatively satisfactory quality to diagnose 'normal' despite intravascular low signal, and "poor" for equivocal diagnosis or non-visualization of the origin or segment of the vessels due to low signal or artifacts which needs catheter angiography. At the level of the carotid bifurcation, it was compared with conventional 2D-TOF MRA image. Overall image quality was also compared visually and quantitatively by measuring signal-to-noise ratios (SNRs) of the ascending aorta, the innominate artery and both common carotid arteries among the three different coils used(CP body array(n=12), CP neck array(n=9), and head-and-neck(n=8). Results : Demonstration of the aortic arch and its major branches was rated as "good" in 55% (16/29) and "fair" in 34%(10/29). At the level of the carotid bifurcation, image quality of turbo CE MRA was same as or better than conventional 2D-TOF MRA in 65% (17/26). Overall image quality and SNR were significantlygreater with CP body array coil than with CP neck array or head-and-neck coil. Conclusions : Turbo CE MRA can be used as a screening exam in the evaluation of the major branches of the aortic arch from their origin to the skull base. Overall imagequality appears to be better with CP body array coil than with CP neck array coil or head-and-neck coil.

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A Study on Vascular Plants, Distribution Status and Management Plans of the Cactus Habitat (No. 429 Natural Monument) in Wolryung-ri, Jeju Island (제주 월령리 선인장군락지(천연기념물 제429호)의 관속식물상, 분포실태, 관리방안에 관한 연구)

  • Lee, Cheol-Ho;Jang, Gye-Hyun;Ryu, Tae-Bok;Choi, Byoung-Ki
    • Journal of the Korean Institute of Traditional Landscape Architecture
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    • v.36 no.1
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    • pp.55-66
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    • 2018
  • The cactus habitat in Jeju Island has a phytogeographically specific distribution in the East Asian region, and forms a unique landscape as the only native cactus (Opuntia ficus-indica (L.) Mill.) habitat in Korea. However, there has been no detailed investigation on the distribution of cacti in the habitat and no investigation on the diversity of the mixed composition of plants in the habitats and their correlation with the distribution of cactus populations. This study attempted to investigate the diversity of vascular plants in the Wollyeong-ri cactus habitat and record the actual distribution and trends of cactus distribution. In addition to the distribution characteristics of cacti, we also discuss the characteristics of species reflecting the characteristics of the habitat among the mixed population of plant species, as well as the biological and environmental factors that threaten the maintenance of cactus populations or require management for preservation of cacti. Considering the phenological character, we conducted the field surveys for flora identification six times between June 2015 and September 2017. The Engler classification system was used for the arrangement sequence and names of plants, and the Korean Plant Names Index was adopted for the Korean name of the species. The study results showed that the Wolryung-ri cactus habitat in Jeju Island has the characteristic physiognomy of an area dominated by cactus. For the vascular plants, a total of 125 taxa were identified, including 53 families, 104 genera, 109 species, 15 varieties and 1 forma. Endangered plants specified by the Ministry of Environment were not found. Two species, Cyrtomium falcatum and Asplenium incisum, were identified as the ferns, and no gymnosperms were found. In addition, 123 taxa of angiosperms, 91 taxa of dicotyledones and 32 taxa of monocotyledons were identified. The distributions of cacti were confirmed in 289 meshes corresponding to 59.3% of the total 487 meshes in the cactus protected area, which showed various coverage distributions ranging from 5% to 95%. Most of the meshes where no cacti were found are coastal areas with exposed basalt rocks where the soil depth has not developed or extremely restricted due to repeated waves, or areas where artificial facilities, grasslands, and observation paths have been constructed. On the other hand, there were 71 lattice points in 14.5% of the total area where the cactus showed 70% or higher dominance. Cacti are randomly distributed in these areas. They have adapted to the microhabitat environment and are found to be opportunistically distributed along the growable locations. Considering that the reproduction of cacti in the habitat is mostly dependent on parthenogenesis, the present distribution seems to reflect the potentially distributable regions of cacti in the habitat. Based on the results of field surveys, a management plan for conservation and protection of the protected areas has been proposed.

Functional Expression of an Anti-GFP Camel Heavy Chain Antibody Fused to Streptavidin (Streptavidin이 융합된 GFP항원 특이적인 VHH 항체의 기능적 발현)

  • Han, Seung Hee;Kim, Jin-Kyoo
    • Journal of Life Science
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    • v.28 no.12
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    • pp.1416-1423
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    • 2018
  • With strong biotin binding affinity ($K_D=10^{-14}M$), the tetrameric feature of streptavidin could be used to increase the antigen binding activity of a camel heavy chain (VHH) antibody through their fusion, here stained with biotinylated horseradish peroxidase and subsequent immunoassays ELISA and Western blot analysis. For this application, we cloned the streptavidin gene amplified from the Streptomyces avidinii chromosome by PCR, and this was fused to the gene of the 8B9 VHH antibody which is specific to green fluorescent protein (GFP) antigens. To express a soluble fusion protein in Escherichia coli, we used the pUC119 plasmid-based expression system which uses the lacZ promoter for induction by IPTG, the pelB leader sequence at the N-terminus for secretion into the periplasmic space, and six polyhistidine tags at the C-terminus for purification of the expressed proteins using an $Ni^+$-NTA-agarose column. Although streptavidin is toxic to E. coli because of its strong biotin binding property, this soluble fusion protein was expressed successfully. In SDS-PAGE, the size of the purified fusion protein was 122.4 kDa in its native condition and 30.6 kDa once denatured by boiling, suggesting the tetramerization of the monomeric subunit by non-covalent association through the streptavidin moiety fusing to the 8B9 VHH antibody. In addition, this fusion protein showed biotin binding activity similar to streptavidin as well as GFP antigen binding activity through both ELISA and Western blot analysis. In conclusion, the protein resulting from the fusion of an 8B9 VHH antibody with streptavidin was successfully expressed and purified as a soluble tetramer in E. coli; it showed both biotin and GFP antigen binding activity suggesting the possible production of a tetrameric and bifunctional VHH antibody.