• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제27권5호
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• pp.373-384
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• 2001

Cellular proliferation is an intricately regulated process mediated by the coordinated interactions of critical growth control genes. Two of these factors in mammalian cells are the p53 and mdm-2 genes. A protein product of the mem-2 oncogene has been recently shown to associate with the protein encoded by the tumor suppressor gene p53. The p53 tumor suppressor protein is stabilized in response to DNA damage and other stress signals and causes the cell to undergo growth arrest or apoptosis, thus preventing the establishment of mutations in future cellular generations. Mutation or loss of p53 is a very common event in tumor progression. It occurs in about 50% of all tumors analysed including of colon, lung, breast and liver. The cellular mdm-2 gene, which has potential transforming activity that can be activated by overexpression, is amplified in a significant percentage of human sarcoma and in other mammalian tumors. Proteins encoded by the mdm-2 gene are able to bind to the p53 protein and, when overexpressed, can inhibit p53's transcriptional activation function, thus mdm-2 can act as a negative regulator of p53 function. Experimental study was performed to observe the relationship between p53 gene mutation and mdm-2 protein expression and apply the results to the clinical activity. 36 golden syrian hamster each weighing $60{\sim}80g$ were used and painted with 0.5% DMBA by 3 times weekly on the right buccal cheek(experimental side) for 6, 8, 10, 12, 14 and 16 weeks. Left buccal cheek(control side) was treated with mineral oil as the same manner to the right side. The hamsters were sacrificed on the 6, 8, 10, 12, 14 & 16 weeks. Normal and tumor tissues from paraffin block were examined for histology and immunohistochemistry observation, and were completely dissected by microdissection and DNA from both tissue were isolated by proteins K/phenol/chloroform extraction. Segments of the hamster p53 exons 5, 6, 7 and 8 were amplified by PCR using the oligonucleotide primers, and then confirmational change was observed by SSCP respectively. The results were as follows : 1. Dysplasia at 6 weeks, carcinoma in situ at 8 weeks and invasive carcinoma from 10 weeks could be observed in experimental groups. 2. p53 mutations were detected in 10 of the 36(28%) and the exons 6(6 of the 10 : 60%) was the most hot spot area among the highy conserved region(exons 5, 6, 7 & 8). 3. Immunohistochemical study confirmed 22 of the 36(61%) of p53 expression involving 10 of p53 mutations. 4. mdm-2 expression of was showed in 3 of the 36(8%) involving 1 of the 22 of p53 expression and 2 of the 14 of p53 non-expression. From the above results, mutation of p53 gene or expression of p53 protein may have the influence of the DMBA induced carcinoma of hamster buccal pouch but the expression of mdm-2 protein may not have relationship with tumorigenesis.

• 식물병연구
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• 제30권1호
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• pp.26-33
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• 2024

2023년 강원특별자치도 철원군의 파프리카 농가 74곳에서 특정 품종의 유묘에 줄기가 잘록해져서 쓰러지는 모잘록병 증상을 보이는 병이 대발생함에 따라 본 연구에서는 모잘록병의 원인을 규명하고자 하였다. 모잘록병으로 의심되는 유묘에서 현미경으로 관찰 시, 초승달 모양의 F. oxysporum의 전형적인 분생포자가 관찰되었다. 이에 따라 F. oxysporum 특이 프라이머를 이용하여 PCR을 수행하여 양성 밴드를 확보하였으며, 염기서열을 분석한 결과 F. oxysporum의 유전자임이 확인되었다. 농가에서 분리한 F. oxysporum의 병원성의 재현성을 확인하기 위해 농가에서 분리한 F. oxysporum 균주를 다른 파프리카 품종의 건전주에 접종하여 모잘록병이 발생함을 확인하였다. 이번 모잘록병의 대발생이 오염된 종자에 의한 것인지, 농가 환경에서 감염되어 확산된 것인지의 여부를 확인하기 위해, 표면살균 처리한 파프리카 종자와 살균하지 않은 무처리 종자를 MS 배지에 각각 치상하여 배양하였다. 그 결과, 무처리 종자배지에서만 분홍색의 전형적인 F. oxysporum의 균사가 확인되었다. 또한, 농가에서 분리한 균주, 농가에서 분리한 균주를 접종하여 얻은 균주 그리고 같은 파프리리카 품종의 제조생산번호가 같은 미개봉 종자에서 분리한 균주가 동일한 F. oxysporum인지를 확인하기 위해 18S rRNA region의 염기서열을 이용하여 분자계통분류학적 분석을 진행한 결과, 결과 3종의 F. oxysporum 균주가 같은 그룹으로 분류되었으며, 기존에 철원군에서 보고된 F. oxysporum과는 다른 그룹임을 확인하였다. 이는 파프리카의 모잘록병 대발생의 원인은 F. oxysporum에 오염된 종자에 의한 것임을 증명하였다.