• Mycobiology
• /
• 제42권4호
• /
• pp.311-316
• /
• 2014

Lichen studies, including biodiversity, phylogenetic relationships, and conservation concerns require definitive species identification, however many lichens can be challenging to identify at the species level. Molecular techniques have shown efficacy in discriminating among lichen taxa, however, obtaining genomic DNA from herbarium and fresh lichen thalli by conventional methods has been difficult, because lichens contain high proteins, polysaccharides, and other complex compounds in their cell walls. Here we report a rapid, easy, and inexpensive protocol for extracting PCR-quality DNA from various lichen species. This method involves the following two steps: first, cell breakage using a beadbeater; and second, extraction, isolation, and precipitation of genomic DNA. The procedure requires approximately 10 mg of lichen thalli and can be completed within 20 min. The obtained DNAs were of sufficient quality and quantity to amplify the internal transcribed spacer region from the fungal and algal lichen components, as well as to sequence the amplified products. In addition, 26 different lichen taxa were tested, resulting in successful PCR products. The results of this study validated the experimental protocols, and clearly demonstrated the efficacy and value of our KCl extraction method applied in the fungal and algal samples.

• 미생물학회지
• /
• 제38권4호
• /
• pp.230-230
• /
• 2002

Class Ⅲ chitin synthases in filamentous fungi are important for hyphal growth and differentiation of several filamentous fungi. A genomic clone containing the full gene encoding Chs4, a class Ⅲ chitin synthase in Penicillium chrysogenum, was cloned by PCR screening and colony hybridization from the genomic library. Nucleotide sequence analysis and transcript mapping of chs4 revealed an open reading frame (ORF) that consisted of 5 exons and 4 introns and encoded a putative protein of 915 amino acids. Nucleotide sequence analysis of the 5′flanking region of the ORF revealed a potential TATA box and several binding sites for transcription activators. The putative transcription initiation site at -716 position was identified by primer extension and the expression of the chs4 during the vegetative growth was confirmed by Northern blot analysis. Amino acid sequence analysis of the Chs4 revealed at least 5 transmembrane helices and several sites for past-transnational modifications. Comparison of the amino acid sequence of Chs4 with those of other fungi showed a close relationship between P chrysogenum and genus Aspergillus.

• 한국유전체학회:학술대회논문집
• /
• 한국유전체학회 2006년도 The 15th Korean Genome Organization Conference KOGO 2006 Annual Meeting
• /
• pp.55-55
• /
• 2006

• 한국당과학회:학술대회논문집
• /
• 한국당과학회 2006년도 제1회 연례학술대회
• /
• pp.21-22
• /
• 2006

• 한국연초학회지
• /
• 제37권1호
• /
• pp.25-33
• /
• 2015

Genome walking is a par ticular application for identifying sequences of unknown genomic regions adjacent to a known region. Many genome walking methods based on polymerase chain reaction (PCR) are available. Even if earlier techniques suffer from low reproducibility, inefficiency, and non-specificity, improved strategies have been developed. In this study, we present an alternative strategy: the genomic DNA is digested with restriction enzymes. After cytosine overhangs at 5' ends, the fragments are ligated to linker adaptor s had guanine overhang at 3' ends. Then nested PCR is performed. The improvements in this strategy focus on two points. The first is the C tailing method using Pfu polymerase instead of the A tailing method based on nontemplate-dependent terminal transferase activity of Taq polymerase. Therefore unintended modification of target DNA can be prevented without A tailing error. The second point is the use of C/G-specific ligation had advantage in the ligation efficiency compared with A/T-specific ligation. Therefore, the C-G linker PCR method increases ligation efficiency between digested genomic DNA and adaptor DNA. As a result, the quantity of target DNA to amplify by PCR is enriched. We successfully used G-C linker PCR to retrieve flanking regions bordering the phophinothricin resistance gene in genetically modified tobacco (GMO).

• Molecules and Cells
• /
• 제19권3호
• /
• pp.303-309
• /
• 2005

The end of eukaryotic genomic DNA is capped by a specialized structure called as "telomere" which consists of the repetitive array of nucleotide sequence, TTAGGG, in humans and mice, and a variety of binding proteins. Telomerase is a ribonucleoprotein (RNP) complex responsible for the elongation of telomeres to maintain the genomic integrity, and is composed of telomerase reverse transcriptase (TERT), telomerase RNA component (TERC), and their associated factors regulating the catalytic activity of telomerase. Although it is now apparent that telomerase protects cells from apoptosis via the maintenance of genomic integrity by stabilizing telomeres, our understanding for the physiological role of telomerase is yet far from completion, and emerging evidence suggests that telomerase has additional extratelomeric roles in mediating cell survival and anti-apoptotic functions against various cytotoxic stresses. Here we summarize and discuss how telomerase and telomeres are involved in mediating cellular protection against apoptosis.

• Journal of Genetic Medicine
• /
• 제18권2호
• /
• pp.121-126
• /
• 2021

Chronic progressive external ophthalmoplegia (CPEO) is a complex slowly progressive mitochondrial disorder characterized by extraocular muscle weakness with or without multisystem involvement. The mainstay of therapy in a patient with CPEO is supportive. However, in moderate cases, surgery might be indicated including surgeries for ptosis and strabismus. In this article, we report a Saudi patient with CPEO due to compound heterozygous variants in the DNA polymerase gamma (POLG) gene c.2246T>C p.(Phe749Ser) and c.1735C>T p.(Arg579Trp), which are classified as pathogenic. Proper diagnosis with genetic testing confirmation is important to guide the management and counsel the patient about the prognosis and the management options. The patient was successfully managed with bilateral frontalis sling and illustrates the importance of surgical intervention to improve vision and cosmetic appearance in patients with CPEO. We emphasize the importance of multidisciplinary care in the management of cases of mitochondriopathy, especially CPEO.

• Asian-Australasian Journal of Animal Sciences
• /
• 제26권12호
• /
• pp.1672-1679
• /
• 2013

Linkage disequilibrium between markers or genetic variants underlying interesting traits affects many genomic methodologies. In many genomic methodologies, the effective population size ($N_e$) is important to assess the genetic diversity of animal populations. In this study, dairy cattle were genotyped using the Illumina BoviveHD Genotyping BeadChips for over 777,000 SNPs located across all autosomes, mitochondria and sex chromosomes, and 70,000 autosomal SNPs were selected randomly for the final analysis. We characterized more accurate linkage disequilibrium in a sample of 96 dairy cattle producing milk in Korea. Estimated linkage disequilibrium was relatively high between closely linked markers (>0.6 at 10 kb) and decreased with increasing distance. Using formulae that related the expected linkage disequilibrium to $N_e$, and assuming a constant actual population size, $N_e$ was estimated to be approximately 122 in this population. Historical $N_e$, calculated assuming linear population growth, was suggestive of a rapid increase $N_e$ over the past 10 generations, and increased slowly thereafter. Additionally, we corrected the genomic relationship structure per chromosome in calculating $r^2$ and estimated $N_e$. The observed $N_e$ based on $r^2$ corrected by genomics relationship structure can be rationalized using current knowledge of the history of the dairy cattle breeds producing milk in Korea.

• 한국축산식품학회지
• /
• 제27권2호
• /
• pp.150-156
• /
• 2007

본 연구는 서로 다른 상염색체에 위치하고 있는 초위성체 유전 표지를 활용한 한국재래돼지 집단의 개체 식별시스템 설정을 위해 수행되었다. 공시재료는 4품종에서 총 446두가 사용되었으며 13종의 좌위에 대한 개체별 유전자형을 분석하였다. 이들 13종에서 출현된 이형접합도는 0.286-0.686였으며 marker 다형성 정보량은 0.399-0.796로 나타났다. 한국 재래돼지 집단에서 나타난 대립 유전자 발현 특성은 다른 대조 품종 집단과 매우 상이한 결과를 나타냈다. S0228좌위는 전체 8종의 대립유전자가 나타난 가운데 다른 3품종에서는 발현이 되지 않은 235 대립유전자가 한국 재래돼지 집단에서만 발현이 되었다. 5종의 초위성체 유전 표지를 활용할 경우 누적 개체 식별력은 99.999%를 나타냈으며 두 마리의 서로 다른 개체가 서로 같은 유전자형을 가질 짝확률은 $0.36{\times}10^{-9}$으로 추정되었다. 따라서 10종의 선정된 유전 표지는 한국재래돼지 집단에서 적정 신뢰도를 제공할 수 있는 개체 식별 시스템을 설정할 수 있을 것으로 생각된다.

• 유전체소식지
• /
• 제4권1호
• /
• pp.13-15
• /
• 2004