• Journal of Microbiology
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• 제45권2호
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• pp.105-112
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• 2007

The objective of this study was to analyze the phylogenetic composition of bacterial community in the soil of an earth-cave (Niu Cave) using a culture-independent molecular approach. 16S rRNA genes were amplified directly from soil DNA with universally conserved and Bacteria-specific rRNA gene primers and cloned. The clone library was screened by restriction fragment length polymorphism (RFLP), and representative rRNA gene sequences were determined. A total of 115 bacterial sequence types were found in 190 analyzed clones. Phylogenetic sequence analyses revealed novel 16S rRNA gene sequence types and a high diversity of putative bacterial community. Members of these bacteria included Proteobacteria (42.6%), Acidobacteria (18.6%), Planctomycetes (9.0 %), Chloroflexi (Green nonsulfur bacteria, 7.5%), Bacteroidetes (2.1%), Gemmatimonadetes (2.7%), Nitrospirae (8.0%), Actinobacteria (High G+C Gram-positive bacteria, 6.4%) and candidate divisions (including the OP3, GN08, and SBR1093, 3.2%). Thirty-five clones were affiliated with bacteria that were related to nitrogen, sulfur, iron or manganese cycles. The comparison of the present data with the data obtained previously from caves based on 16S rRNA gene analysis revealed similarities in the bacterial community components, especially in the high abundance of Proteobacteria and Acidobacteria. Furthermore, this study provided the novel evidence for presence of Gemmatimonadetes, Nitrosomonadales, Oceanospirillales, and Rubrobacterales in a karstic hypogean environment.

• 원예과학기술지
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• 제33권3호
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• pp.396-402
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• 2015

This study identified a new male-sterile germplasm of Chinese jujube, named male-sterile No. 2 (JMS2), and achieved controlled hybridization using that germplasm as the female parent. The anthers of JMS2 before flower bud opening became shrunken, dingy yellow and much smaller than normal ones, and they changed to brown after anthesis. No pollen was observed in anthers of JMS2 and its male-sterile trait remained stable over different years. A total of 1,642 fruits were obtained from ten intra- and interspecific cross combinations via controlled hybridization from 2008 to 2012 using JMS2 as the female parent. Of those, 27.3% produced seeds, on average (0-72.6%). The rate of fruit with seed (RFS) was significantly different between cross combinations or years. Compared to other cross combinations, the RFS in the combination of JMS2 ${\times}$ 'Xingguang' (a Chinese jujube cultivar with high resistance to jujube witches' broom disease) and JMS2 ${\times}$ 'Xing16' (a sour jujube genotype) remained high in different years and reached means of 48.7 and 58.1%, respectively. Finally, 150 plantlets were regenerated from immature embryos, and 51 of them were randomly selected and identified to be authentic hybrids using amplified fragment length polymorphism (AFLP) markers. This is the first report of hybrids obtained from a cross between Chinese jujube and sour jujube.

• 한국작물학회지
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• 제45권2호
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• pp.123-127
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• 2000

The objective of this study was to develop a linkage map of soybean under the genetic background of Korean soybean. A set of 89 F/sub 5/ lines was developed from a cross between 'Pureunkong', which was released for soy-bean sprout, and 'Jinpumkong 2', which had no beany taste in seed due to lack of lipoxygenase 1, 2, and 3. A linkage map was constructed for this population with a set of 113 genetic markers including 7 restriction fragment length polymorphism (RFLP) markers, 79 randomly amplified polymorphic DNA (RAPD) markers, 24 simple sequence repeat(SSR) markers, and 3 morphological markers. The map defined approximately 807.4 cM of the soybean genome comprising 25 linkage groups with 98 polymorphic markers. Fifteen markers remained unlinked. Seventeen linkage groups identified here could be assigned to the respective 13 linkage groups in the USDA soybean genetic map. RFLP and SSR markers segregated at only single genetic loci. Fourteen of the 25 linkage groups contained at least one SSR marker locus. Map positions of most of the SSR loci and their linkages with RFLP markers were consistent with previous reports of the USDA soybean linkage groups. For RAPD, banding patterns of 13 decamer primers showed independent segregations at two or more marker loci for each primer. Only the segregation at op Y07 locus was expressed with codominant manner among all RAPD loci. As the soybean genetic map in our study is more updated, molecular approaches of agronomically important genes would be useful to improve Korean soybean improvement.

• The Plant Pathology Journal
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• 제25권1호
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• pp.13-20
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• 2009

Ninety isolates of Colletotrichum species from new persimmon tree twigs and 50 isolates from pepper plant fruits were isolated via single-spore isolation. Of the 140 isolates, 26 were examined for mycelial growth, carbendazim sensitivity, and ITS sequence. Four of the isolates from the persimmon trees, which were cultivated exclusively in an orchard, showed fast mycelial growth and sensitivity to carbendazim, while five of the pepper isolates showed slower mycelial growth and were resistant to the fungicide. However, 17 isolates from persimmon trees cultivated with pepper plants in the same orchard showed slow mycelial growth like the pepper isolates and they were sensitive to carbendazim like the persimmon isolates. ITS sequence analysis of these 27 isolates led to the identification of the 22 persimmon isolates as C. gloeosporioides and the five pepper isolates as C. acutatum. PCR with species-specific primers confirmed that the 90 isolates from persimmon were C. gloeosporioides whereas the 50 isolates from pepper were C. acutatum. The 90 persimmon isolates of C. gloeosporioides and 50 pepper isolates of C. acutatum were compared by a wound inoculation test to determine their capacity for host cross-infection. All of the C. acutatum isolates from pepper caused typical symptoms of anthracnose on the fruits of pepper plants and twigs of persimmon; they differed from the C. gloeosporioides isolates from persimmon, more than 90% of which were able to infect only persimmon. Amplified fragment length polymorphism analysis revealed the existence of two groups (C. gloeosporioides and C. acutatum isolates group). At 80% genetic similarity, the C. gloeosporioides group was defined within four clusters, while the C. acutatum group was within three clusters. However, these clusterings were unrelated with the virulence of Colletotrichum species against pepper fruits.

• Journal of Plant Biotechnology
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• 제7권2호
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• pp.81-86
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• 2005

Panax ginseng is one of the most important medicinal plants in the world. The international trade of ginseng is increasing yearly. The disguise of Chinese and American ginseng into Korean ginseng became a problem in recent years in abroad and Korea. An effective method to authenticate the Korean Panax ginseng from others at a DNA level is necessary for the healthy development of the ginseng market. Amplified fragment length polymorphism (AFLP) analysis was applied to develop a method for the identification of Korean ginseng between Chinese ginseng and American ginseng. It is very difficult to detect the different polymorphic bands among Korean field cultivated ginseng, and between field and wild-cultivated ginseng. The genetic distance coefficient by AFLP analysis between field- and wild cultivated Korean ginseng was very low, 0.056. Whereas, polymorphic bands between Korean and Chinese wild-cultivated ginseng was significantly different. The genetic distance coefficient between wild-cultivated Korean and Chinese ginseng was 0.149. The genetic distance coefficients between the P. ginseng and P. quinquefolius were ranging from 0.626 to 0.666. These results support that the AFLP analysis could be applied to authenticate Korean P. ginseng from others Chinese P. ginseng and American ginseng (P. quinquefolius).

• Parasites, Hosts and Diseases
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• 제35권4호
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• pp.271-276
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• 1997

메타고니무스속 흡충의 형태학적인 차이점은 잘 알려져 있으나 이러한 미세한 형태학적 차이로 종을 분류할 수 있을 지에 대해서는 의문시되어 왔다. 이 연구는 비교적 유전자 염기서열이 잘 보존되어 있 어 종간 또는 strain간의 차이를 밝힐 수 있는 리보솜리보핵산 유전자 중 ITSI 유전자와 사립체 COI 유전자를 중합효소반응으로 증폭시킨 후 제한효소로 소화시켜 나타나는 밴드의 차이를 관찰하였다 요 코가와흡충 (M. yokogawai)의 피 낭유충은 삼척산 은어에서 , 미야타흡충 (Metagonim Miyata type) 은 충주산 피라미에서, 타카하시홉충 (M. tnkqhqsrii)은 충주산 붕어에서 분리하여 사용하였다. 세 종류 충체에서 얻은 ml 유전자 증폭산물은 제한효소 Rsc I, Ak I 및 Msp I에 의해 서로 다른 크기의 밴드 로 소화되었다. 세 종류 충체의 사립체 COI 유전자 증폭산물도 Rsc I과 AIu I에 의해 서로 다른 양상으로 잘라졌다. 추정 유전자 차이 (estimated genetic divergence)는 미야타홉충과 요코가와흡충이 0.034880, 요코가와흡충과 타카하시홉충이 0.018179, 미야타흡충과 타카하시흡충이 0.028098 이었다. 이 결과로 보면 미야타흡충은 별개의 종으로 볼 수 있으며,다른 충체보다 이른 시기에 진화하였음 을 알 수 있다.

• 한국식품위생안전성학회지
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• 제15권2호
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• pp.128-136
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• 2000

수입 냉동어패류에서 24주의 Vibiro 균주를 순수 분리하여 그들의 생리, 생화학적 특성에 따라 동정한 결과 V. cholerae non-O1과 V. diazotrophicus가 각 2주, V. hollisae 1주, V. natriegens 5주, V. fluvialis 8주, V. nereis 4주인 것으로 밝혀졌고 2주는 Vibirio속 균주가 아닌 것으로 나타났다. 이 균주들을 API-2OE kit로 동정한 결과는 위의 결과와 매우 달라, V. cholerae로 동정된 2주와 fluvialis로 동정된 5균주의 경우에만 결과가 일치하였고 나머지는 Vibrio속이 아닌 것으로 분석되었다. 식품류에서 V. cholerae가 검출된 점에 주목하여 이 분리균들의 동정을 보다 정확히 하기 위하여 분리균들의 165 rRNA를 증폭시켜 이들의 RFLP를 분석하였다. 그 결과, r cholerae로 동정된 두 균주는 동일한 RFLP양상을 갖고 있었으며, V. cholerae보다는 V. proteolyticus의 RFLP에 보다 가까운 것으로 나타났다. 따라서 식품류의 병원균 검색을 보다 정확하고 신속하게 할 수 있도록 효율적인 검색 방법의 개발이 시급하다고 하겠다.

• 한국양식학회지
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• 제20권4호
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• pp.255-259
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• 2007

우리나라 나로도, 태안, 영광 및 중국 보하이만에서 채집된 4개의 대하(Fenneropenaeus chinensis) 자연산 집단에 대한 유전학적 다양성 및 근연 관계를 AFLP 지문 분석을 통해 조사하였다. 5종의 primer 조합형을 이용한 AFLP 분석에서 각 집단으로 부터 $251{\sim}254$개의 bands를 얻어 분석한 결과, 집단내 다형 band의 출현 빈도는 4개 집단에서 $27.1{\sim}28.1%$로 유사하게 나타났고 이형접합율($0.1177{\sim}0.1288$)및 유전적 다양도($0.1099{\sim}0.1194$) 역시 4개 집단에서 동일한 수준을 보였다. Pairwise distance, 유전적 분화도(Fst index) 및 유전적 상동성 분석 역시 유사한 결과를 나타내어 본 연구에서 분석한 4개 대하 집단은 유전적으로 매우 밀접한 근연 관계를 나타내었고 특정 집단의 유전적 분화는 없는 것으로 판단되었다.

• Asian-Australasian Journal of Animal Sciences
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• 제29권4호
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• pp.564-570
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• 2016

The Sal-like 1 gene (Sall1) is essential for kidney development, and mutations in this gene result in abnormalities in the kidneys. Mice lacking Sall1 show agenesis or severe dysgenesis of the kidneys. In a recent study, blastocyst complementation was used to develop mice and pigs with exogenic organs. In the present study, transcription activator-like effector nuclease (TALEN)-mediated homologous recombination was used to produce Sall1-knockout porcine fibroblasts for developing knockout pigs. The vector targeting the Sall1 locus included a 5.5-kb 5' arm, 1.8-kb 3' arm, and a neomycin resistance gene as a positive selection marker. The knockout vector and TALEN were introduced into porcine fibroblasts by electroporation. Antibiotic selection was performed over 11 days by using $300{\mu}g/mL$ G418. DNA of cells from G418-resistant colonies was amplified using polymerase chain reaction (PCR) to confirm the presence of fragments corresponding to the 3' and 5' arms of Sall1. Further, mono- and bi-allelic knockout cells were isolated and analyzed using PCR-restriction fragment length polymorphism. The results of our study indicated that TALEN-mediated homologous recombination induced bi-allelic knockout of the endogenous gene.

• Journal of Oral Medicine and Pain
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• 제21권2호
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• pp.351-367
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• 1996

In the field Of individual identification in forensic Science, if the body is charred, it is sometimes impossible to identify the morphologic changes and charred tissue such as blood, muscle and bone can not be identified by forensic microbiologic method such as DNA typing. So the author used the characteristics of teeth which is relatively firm compare to other organs and stable to external environment such as heat and also possess cells needed for the DNA typing. The author conducted the experiment on teeth to detect DNA related to individual identification regarding to temperature in which other charredorgans can not be detected. The experiment was done on 64 extracted third molars consisted of unheated ones, and heated teeth to $100^{\circ}C$, $150^{\circ}C$, $200^{\circ}C$ for 45 min, 90 min, and 120 min respectively and to $250^{\circ}C$ for 45 min. DNA was extracted from each tooth and amplified fragment length polymorphism procedure(AMP-FLPs) using polymerase chain reaction(PCR) was applied and observed for the matching DNA in HumTH01 and HumCD4 locus and the followings Are the results : 1. It was able to detect matching DNA in HumTH01 and HumCD4 locus in every teeth which no heating has been done. 2. It was able to detect matching DNA in HumTH01 and HumCD4 locus in every teeth heated to $100^{\circ}C$ for 45, 90 and 120 min. 3. It was able to detect matching DNA in HumTH01 and HumCD4 locus in teeth heated to $l00^{\circ}C$, $200^{\circ}C$ for 45, 90, 120 min. 4. It was impossible to detect matching DNA in HumTH01 and HumCD4 locus in teeth heated to $250^{\circ}C$. So, it is possible to extract DNA from teeth that otherwise can not be extracted from other organs in the charred body and it can be concluded that teeth are highly reliable and applicatable as forensic odontology for individual identification.