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Identification and Characterization of Three Isolates of Cucumber mosaic virus Isolated from Weed Hosts (잡초에서 분리한 3종 Cucumber mosaic virus의 동정과 특성)

  • Lee, Hyeok-Geun;Kim, Sung-Ryul;Jeon, Yong-Woon;Kwon, Soon-Bae;Ryu, Ki-Hyun;Choi, Jang-Kyung
    • Research in Plant Disease
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    • v.14 no.1
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    • pp.15-20
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    • 2008
  • Three isolates of Cucumber mosaic virus (CMV) were isolated from weed hosts showing typical mosaic symptoms, and some properties of the viruses were investigated. CMV isolates, designated as Is-CMV, Jd-CMV and Pla-CMV from Isodon inflexus, Jeffersonia dubia and Phryma leptostachya var. asiatica, respectively, were identified and characterized by biological reaction in several host plants, serological property, dsRNA analysis, reverse transcription-polymerase chain reaction (RT-PCR), restriction fragment-length polymorphism (RFLP). All isolates systemically infected in Nicotiana benthamiana, Cucurbita pepo cv. Black beauty and Cucumis sativus, and did not reveal any differences in these host plants between the isolates. However, remarkable difference in the symptoms was found between the CMVs in Capsicum annuum. Is-CMV induced an asymptomatic symptoms, while Jd-CMV and Pla-CMV produced severe mosaic symptoms in C. annuum plants. In dsRNA analysis, all isolates revealed four major bands with estimated molecular size of 3.4, 3.2, 2.1 and 1.0 kbp. The cDNAs of coat protein gene of the isolates were amplified by RT-PCR using a genus-specific single pair primers that designed to amplify a DNA fragment of approximately ranging from 938 to 966 bp. By restriction mapping analysis using RFLP of the RT-PCR products as well as by serological properties of gel diffusion test, the CMV isolates belong to a typical members of CMV subgroup IA. This is the first report on the occurrence of CMV in the three weed hosts.

Genomic analysis of Mycobacterium fortuitum by pulsed-field gel electrophoresis (Pulsed-field Gel Electrophoresis를 이용한 Mycobacterium fortuitum의 유전형 분석)

  • Lee, Tae-Yoon;Do, In-A;Kim, Sung-Kwang
    • Journal of Yeungnam Medical Science
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    • v.12 no.2
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    • pp.366-385
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    • 1995
  • Epidemiological studies are important in both the prevention and treatment of mycobacterial infections. This study was initiated to establish the pulsed-field gel electrophoresis (PFGE) method, which are not yet extensively studied. The most apprpriate restriction endonucleases included DraI, AsnI, and XbaI. The optimal PFGE condition was different according to the enzymes used. Two stage PFGE was performed, in case of DraI first stage was performed with 10 seconds of initial pulse and 15 seconds of final pulse, while the second stage was performed with 60 seconds of initial pulse and 70 seconds of final pulse. The electrophoresis time for DraI-PFGE was 14 hours for each stage. Electrophoresis was performed for 22 hours, in case of XbaI, with 3 seconds of initial pulse and 12 seconds of final pulse. Electrophoresis was performed for 22 hours, in case of AsnI, with 5 seconds of initial pulse and 25 seconds of final pulse. In all cases the voltage of the electrophoresis was maintained constantly at 200 voltage. Standard mycobacterial strains, which included Mycobacterium bovis BCG, M. tuberculosis, and M. fortuitum, could not be differentiated by PFGE analysis. PFGE analysis was performed to differentiate 9 clinically isolated M. fortuitum strains using AsnI. All M. fortuitum strains showed different genotypes except 2 strains. Cluster analysis divided M. fortuitum strains into 2 large groups. PFGE analysis was performed to further differentiate M. fortuitum isolates using XbaI. The undifferentiated 2 M. fortuitum strains showed different PFGE patterns with Xba I. Cluster analysis of the XbaI-PFGE patterns showed more complex grouping than AsnI-PFGE patterns, which showed that XbaI-PFGE analysis was better than AsnI-PFGE in M. fortuitum genotyping. The top dissimilarity values of AsnI-PFGE and XbaI-PFGE were 0.74 and 0.75, respectively. This value was higher than that of arbitrarily primed polymerase chain reaction (AP-PCR) analysis and lower than that of restriction fragment length polymorphism (RFLP) analysis. This suggested that PFGE can be used as a supportive or alternative genotyping method to RFLP analysis.

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A Study on the Characteristics of Seoul Olympic Organizing Committee's Official Documents (서울올림픽대회 조직위원회 공문서의 성격에 관한 연구)

  • Cheon, Ho-Jun
    • The Korean Journal of Archival Studies
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    • no.24
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    • pp.113-171
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    • 2010
  • The purpose of this study was to examine the characteristics of Seoul Olympic Organizing Committee's official documents. To conduct this work, the fundamental of producing archives were examined by analyzing structure and management of Seoul Olympic Organizing Committee and structure of official document production. After all, simultaneous and synthesis characteristics of Seoul Olympic Organizing Committee's official documents were presented through overall analysis of production fundamental and relationship between their management and remained archives. The result of this study are as follows. Firstly, The Organizing Committee had bicameral organizational structure and matrix organizational format consisting of functional department and project department. Indicating the institutions and development phase of decision making in the committee, most of institutions were in name only. Also, there were many problems occurred in the procedure of decision making since the president of committee exercised all of the authorities. Secondly, It was found that existing official documents of the committee were partial and caused fragment phenomenon and severe situations because of unsystematic archival management department and regulations. Moreover, as the result of investigating production procedure and management of official documents, procedure of production, distribution, preservation and abolition of them were specifically verified. Thirdly, It was verified that the official documents were abolished arbitrarily because of unsystematic archival management department and insufficient regulations. For the actual condition of management, filing or description activity which is essential measure for using and utilizing the official documents has not been conducted yet. Based on these facts, the characteristics of Seoul Olympic Organizing Committee's official documents can be referred as follows. The official archives of the committee have multiplicity of the origin and severe fragment phenomenon damaging the origin and the elementary substance of the archives. Also, the format of existing archives was unbalance. Besides, there was not enough related research since they were in adverse situation to utilize them as the archives which are not assessed or not arranged. Thus, it was hard to grasp the utility value at present and future, and was also limited for usage object.

Treatment of Distal Tibial Spiral Fractures Combined with Posterior Malleolar Fractures (후과 골절이 동반된 경골 원위부 나선상 골절의 치료)

  • Kim, Young Sung;Lee, Ho Min;Kim, Jong Pil;Chung, Phil Hyun;Park, Soon Young
    • Journal of the Korean Orthopaedic Association
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    • v.56 no.4
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    • pp.317-325
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    • 2021
  • Purpose: This study compared the functional and radiologic outcomes of intramedullary nailing (IMN) and minimally invasive plate osteosynthesis (MIPO) for tibia fractures in distal tibial spiral fractures combined with posterior malleolar fractures, as well as the functional and radiologic outcomes with and without fixation for posterior malleolar fractures. Materials and Methods: From January 2010 to December 2018 the radiological and clinical outcomes of 30 skeletally mature patients with tibial spiral fractures (AO Foundation/Orthopaedic Trauma Association classification 42-A1, B1, C1) combined with posterior malleolar fractures were analyzed. Sixteen patients were treated with IMN, and 14 patients were treated with MIPO. Depending on the surgical methods, the radiologic and clinical outcomes were compared by evaluating the bone union time, postoperative alignment, postoperative displacement of the posterior malleolar fragment, and American Orthopaedic Foot and Ankle Society (AOFAS) score. Moreover, the functional and clinical outcomes with and without fixation for posterior malleolar fractures were compared. Results: The mean bone union time was 21.8 weeks in the IMN group and 23.1 weeks in the MIPO group (p=0.500). At the final follow up, the mean alignment was coronal angulation of 1.8°, sagittal angulation of 1.6° in the IMN group and coronal angulation of 1.2° and sagittal angulation of 1.7° in the MIPO group (conoral angulation: p=0.131, sagittal angulation: p=0.850). The postoperative and final radiologic evaluation showed no displacement of the posterior malleolar fragment and excellent joint congruity in all cases. At the final follow-up, the mean AOFAS score was 88.0 on average in the IMN group and 87.6 on average in the MIPO group (p=0.905). The ankle range of motion and AOFAS score were similar in the fixation group and no fixation group for posterior malleolar fractures. Conclusion: Both IMN and MIPO for tibial spiral fractures combined with posterior malleolar fractures result in satisfactory radiological and clinical outcomes.

Comparison of the Outcomes according to the Injury Type of the Short Radiolunate Ligament in Fracture-Dislocation of the Radiocarpal Joint (요수근 관절의 골절-탈구에서 단요월상인대의 손상 형태에 따른 치료 결과의 비교)

  • Heo, Youn Moo;Kim, Tae Gyun;Song, Jae Hwang;Jang, Min Gu;Lee, Seok Won
    • Journal of the Korean Orthopaedic Association
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    • v.56 no.1
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    • pp.51-60
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    • 2021
  • Purpose: Radiocarpal dislocation (RCD), which is caused by high-energy trauma, often involves radial styloid fractures and short radiolunate ligament (SRLL) injuries. Although SRLL injuries may occur as a simple rupture at the attachment site of radius, it may occur with a relatively large avulsed-fragment in the volar rim of the lunate facet of the radius. This study aimed to differentiate the injury type of SRLL and assess the differences in the treatment results depending on the treatment methods that have been applied in RCD with radial styloid fractures. Materials and Methods: Eighteen patients managed surgically with RCD were enrolled in this study. The patients were classified as Group 1 and Group 2 by using the Dumontier method. In this study, Group 2 was subdivided into 2A (purely ligamentous or small avulsion fracture of the volar rim of lunate facet) and 2B (large avulsed-fragment enough to internal fixation) according to the injury type of SRLL. Groups 2A and 2B were treated with direct repair and screw fixation, respectively. Pain, range of motion of the wrist joint, grip strength, and complications on final radiographs were examined. The outcomes were evaluated using patient-rated wrist evaluation (PRWE), and modified Mayo wrist score (MMWS). Results: All patients were Group 2 (six and twelve patients in 2A and 2B, respectively). The mean flexion to extension arch recovered 79%,and the mean grip strength was 72.9% of the uninjured side. Group 2A showed better recovery in extension, flexion and pronation than Group 2B, but there was no difference in radial deviation, ulnar deviation, supination, grip strength and pain. No differences in the PRWE and MMWS were observed between two groups. Complications included traumatic arthritis in seven patients and residual instability in five patients. Conclusion: When the SRLL was injured, the involvement of a large avulsion fracture on the anterior plane of the radiolunate did not affect the test results. On the other hand, it should be observed cautiously because avulsion fractures tend to disturb the joint's reduction through rotation or displacement. In addition, anatomical reduction and sturdy internal fixation are important for restoring the function of the SRLL.

Identification of Domesticated Silkworm Varieties Using a Whole Genome Single Nucleotide Polymorphisms-based Decision Tree (전장유전체 SNP 기반 decision tree를 이용한 누에 품종 판별)

  • Park, Jong Woo;Park, Jeong Sun;Jeong, Chan Young;Kwon, Hyeok Gyu;Kang, Sang Kuk;Kim, Seong-Wan;Kim, Nam-Suk;Kim, Kee Young;Kim, Iksoo
    • Journal of Life Science
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    • v.32 no.12
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    • pp.947-955
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    • 2022
  • Silkworms, which have recently shown promise as functional health foods, show functional differences between varieties; therefore, the need for variety identification is emerging. In this study, we analyzed the whole silkworm genome to identify 10 unique silkworm varieties (Baekhwang, Baekok, Daebaek, Daebak, Daehwang, Goldensilk, Hansaeng, Joohwang, Kumkang, and Kumok) using single nucleotide polymorphisms (SNP) present in the genome as biomarkers. In addition, nine SNPs were selected to discriminate between varieties by selecting SNPs specific to each variety. We subsequently created a decision tree capable of cross-verifying each variety and classifying the varieties through sequential analysis. Restriction fragment length polymorphism (RFLP) was used for SNP867 and SNP9183 to differentiate between the varieties of Daehwang and Goldensilk and between Kumkang and Daebak, respectively. A tetra-primer amplification refractory (T-ARMS) mutation was used to analyze the remaining SNPs. As a result, we could isolate the same group or select an individual variety using the nine unique SNPs from SNP780 to SNP9183. Furthermore, nucleotide sequence analysis for the region confirmed that the alleles were identical. In conclusion, our results show that combining SNP analysis of the whole silkworm genome with the decision tree is of high value as a discriminative marker for classifying silkworm varieties.

Diagnostic Value of CYFRA 21-1 Measurement in Fine-Needle Aspiration Washouts for Detection of Axillary Recurrence in Postoperative Breast Cancer Patients (유방암 수술 후 액와림프절 재발 진단에 있어서의 미세침세척액 CYFRA 21-1의 진단적 가치)

  • So Yeon Won;Eun-Kyung Kim;Hee Jung Moon;Jung Hyun Yoon;Vivian Youngjean Park;Min Jung Kim
    • Journal of the Korean Society of Radiology
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    • v.81 no.1
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    • pp.147-156
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    • 2020
  • Purpose The objective of this study was to evaluate the diagnostic value and threshold levels of cytokeratin fragment 21-1 (CYFRA 21-1) in fine-needle aspiration (FNA) washouts for detection of lymph node (LN) recurrence in postoperative breast cancer patients. Materials and Methods FNA cytological assessments and CYFRA 21-1 measurement in FNA washouts were performed for 64 axillary LNs suspicious for recurrence in 64 post-operative breast cancer patients. Final diagnosis was made on the basis of FNA cytology and follow-up data over at least 2 years. The concentration of CYFRA 21-1 was compared between recurrent LNs and benign LNs. Diagnostic performance and cut-off value were evaluated using a receiver operating characteristic curve. Results Regardless of the non-diagnostic results, the median concentration of CYFRA 21-1 in recurrent LNs was significantly higher than that in benign LNs (p < 0.001). The optimal diagnostic cut-off value was 1.6 ng/mL. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of CYFRA 21-1 for LN recurrence were 90.9%, 100%, 100%, 98.1%, and 98.4%, respectively. Conclusion Measurement of CYFRA 21-1 concentration from ultrasound-guided FNA biopsy aspirates showed excellent diagnostic performance with a cut-off value of 1.6 ng/mL. These results indicate that measurement of CYFRA 21-1 concentration in FNA washouts is useful for the diagnosis of axillary LN recurrence in post-operative breast cancer patients.

Establishment and Application of Molecular Genetic Techniques for Preimplantation Genetic Diagnosis of Osteogenesis Imperfecta (골형성부전증의 착상전 유전진단을 위한 분자유전학적 방법의 조건 확립과 적용)

  • Kim, Min-Jee;Lee, Hyoung-Song;Choi, Hye-Won;Lim, Chun-Kyu;Cho, Jae-Won;Kim, Jin-Young;Song, In-Ok;Kang, Inn-Soo
    • Clinical and Experimental Reproductive Medicine
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    • v.35 no.2
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    • pp.99-110
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    • 2008
  • Objectives: Preimplantation genetic diagnosis (PGD) has become an assisted reproductive technique for couples carrying genetic conditions that may affect their offspring. Osteogenesis imperfecta (OI) is an autosomal dominant disorder of connective tissue characterized by bone fragility and low bone mass. At least 95% of cases are caused by dominant mutations in the COL1A1 or COL1A2. In this study, we report on our experience clinical outcomes with 5 PGD cycles for OI in two couples. Methods: Before clinical PGD, we assessed the amplification rate and allele drop-out (ADO) rate of alkaline lysis and nested PCR protocol using heterozygous patient's single lymphocytes in the pre-clinical diagnostic tests for OI. We performed 5 cycles of PGD for OI by nested PCR for the causative mutation loci, COL1A1 c.2452G>A and c.3226G>A, in case 1 and case 2, respectively. The PCR products were analyzed by agarose gel electrophoresis, restriction fragment length polymorphism (RFLP) analysis with HaeIII restriction enzyme in the case 1 and direct DNA sequencing. Results: We confirmed the causative mutation loci, COL1A1 c.2452G>A in case 1 and c.3226G>A in case 2. In the pre-clinical tests, the amplification rate was 94.2% and ADO rate was 22.5% in case 1, while 98.1% and 1.9% in case 2, respectively. In case 1, a total of 34 embryos were analyzed and 31 embryos (91.2%) were successfully diagnosed in 3 PGD cycles. Eight out of 19 embryos diagnosed as unaffected embryos were transferred in all 3 cycles, and in the third cycle, pregnancy was achieved and a healthy baby was delivered without any complications in July, 2005. In case 2, all 19 embryos (100.0%) were successfully diagnosed and 4 out of 11 unaffected embryos were transferred in 2 cycles. Pregnancy was achieved in the second cycle and the healthy baby was delivered in March, 2008. The causative locus was confirmed as a normal by amniocentesis and postnatal diagnosis. Conclusions: To our knowledge, these two cases are the first successful PGD for OI in Korea. Our experience provides a further demonstration that PGD is a reliable and effective clinical techniques and a useful option for many couples with a high risk of transmitting a genetic disease.

Spawning Patterns of Three Bitterling Fishes (Pisces: Acheilognathinae) in Relation to the Shell Size of Host Mussels (Unio douglasiae sinuolatus) (납자루아과(Pisces: Acheilognathinae) 담수어류 3종의 숙주조개(작은말조개; Unio douglasiae sinuolatus) 크기에 대한 산란양상)

  • Choi, Hee-kyu;Lee, Hyuk Je
    • Korean Journal of Environment and Ecology
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    • v.33 no.2
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    • pp.202-215
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    • 2019
  • This study was conducted to investigate the spawning preference of the Acheilognathinae fishes in relation to the shell size of host mussels after identifying the species of eggs and fries in the host mussel using our recently developed RFLP (Restriction Fragment Length Polymorphism) molecular marker at four sites [Hongcheon Naechoncheon (HN) and Deokchicheon (HD) from the North Han River basin and Jeongseon Goljicheon (JG) and Joyanggang (JJ) from the South Han River] in South Korea during May in each year between 2015 and 2018. The Acheilognathinae fish observed in the studied sites included one species (Acheilognathus signifer) in HN and JG, three species (Rhodeus uyekii, A. signifer, and Acheilognathus yamatsutae) in HD, and two species (A. signifer and Acheilognathus yamatsutae) in JJ, and we collected 982 host mussels (Unio douglasiae sinuolatus) that inhabited in all four sites. Using the RFLP molecular marker, we confirmed 46 eggs and fry of the Acheilognathinae fish (454 A. signifer, 43 Acheilognathus yamatsutae, and 149 Acheilognathus yamatsutae) in Unio douglasiae sinuolatus (N=163; 16.6%). We compare the average shell length, shell height, and shell width of mussels with [presence] eggs/fry and mussels without [absence] eggs/fry to examine the spawning preference according to the size of host mussels in each site. The results show that the shell length (1.98 mm), shell height (0.85 mm), and shell width (0.73 mm) of mussels with the eggs/fry were significantly larger (Mann-Whitney U test, P=0.002; difference=1.98 mm) than those of mussel without eggs/fry in HD where three species cohabitated. Although the shell length, shell height, and shell width of mussels with the eggs/fry were larger also in the other three sites, the difference was not statistically significant. In addition, we analyzed the mean number of spawned eggs and fry of each species and found $9.31{\pm}5.94$ R. uyekii, $2.86{\pm}2.45$ A.signifer, and $2.50{\pm}1.32$ A. yamatsutae. R. uyekii spawned 6.45-6.81 more eggs than A.signifer and A. yamatsutae on average per mussel, and it was statistically significant (Kruskal-Wallis test, P < 0.001). These findings indicate that the three species of Acheilognathinae fish tend to prefer larger mussels as their spawning hosts, and this tendency increases when the number of cohabitating bitterling fish species increases. Moreover, A.signifer and A. yamatsutae spawned a smaller number of eggs evenly in more host mussels while R. uyekii spawned many eggs on relatively fewer mussels. We found mussels (N=4) having the eggs/fry of two coexisting species, A. signifier and A. yamatsutae in HD and JJ where more than two bitterling fish species occurred. It suggests the interspecific competition taking place between the Acheilognathinae fishes for utilizing the same resource of mussels for spawning when two or more species cohabitate. This study is expected help to understand better the spawning patterns and reproductive ecology of the Acheilognathinae fishes, which will provide insightful information for advancing our understanding of their ecological relationships - mutualism or host-parasitism - with host mussels.

Protective Immune Response of Bacterially-Derived Recombinant FaeG in Piglets

  • Yahong, Huang;Liang, Wanqi;Pan, Aihu;Zhou, Zhiai;Wang, Qiang;Huang, Cheng;Chen, Jianxiu;Zhang, Dabing
    • Journal of Microbiology
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    • v.44 no.5
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    • pp.548-555
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    • 2006
  • FaeG is the key factor in the infection process of K88ad enterotoxigenic Escherichia coli (ETEC) fimbrial adhesin. In an attempt to determine the possibility of expressing recombinant FaeG with immunogenicity for a new safe and high-production vaccine in E. coli, we constructed the recombinant strain, BL21 (DE3+K88), which harbors an expression vector with a DNA fragment of faeG, without a signal peptide. Results of 15% SDS-polyacrylamide slab gel analysis showed that FaeG can be stably over-expressed in BL21 (DE3+K88) as inclusion bodies without FaeE. Immunoglobulin G (IgG) and M (IgM) responses in pregnant pigs, with boost injections of the purified recombinant FaeG, were detected 4 weeks later in the sera and colostrum. An in vitro villius-adhesion assay verified that the elicited antibodies in the sera of vaccinated pigs were capable of preventing the adhesion of K88ad ETEC to porcine intestinal receptors. The protective effect on the mortality rates of suckling piglets born to vaccinated mothers was also observed one week after oral challenge with the virulent ETEC strain, $C_{83907}$ (K88ad, $CT^+,\;ST^+$). The results of this study proved that the adhesin of proteinaceous bacterial fimbriae or pili could be overexpressed in engineered E. coli strains, with protective immune responses to the pathogen.