PCR PRIMERS FOR THE AMPLIFICATION OF MITOCHONDRIAL SMALL SUBUNIT RIBOSOMAL DNA OF LICHEN-FORMING ASCOMYCETES |
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| Affiliation: | 1. Faculty of Agriculture, Kyushu University Graduate School, 6-10-1 Hakozaki, Higashi-ku, Fukuoka 812-8581, Japan;2. Institute for Protein Research, Osaka University, Suita 565-0871, Japan;3. Department of Molecular Behavioral Biology, Osaka Bioscience Institute, 6-2-4 Furuedai, Suita, Osaka 565-0874, Japan;1. Department of Internal Medicine, University of Pittsburgh Medical Center, Pittsburgh, USA;2. Division of Gastroenterology, Hepatology & Nutrition, Department of Internal Medicine, University of Pittsburgh Medical Center, Pittsburgh, USA;1. Department of Plant Pathology and Microbiology, National Taiwan University, No. 1, Sec. 4, Roosevelt Road, Taipei, 106, Taiwan;2. Taichung District Agricultural Research and Extension Station, No. 370, Song Hwai Road, Dacun Township, Changhua, 515, Taiwan;3. Department of Plant Pathology, National Chung Hsing University, No. 145, Xingda Road, Taichung, 402, Taiwan;4. Institute of Plant Physiology and Genetics, Bulgarian Academy of Sciences, Acad. G. Bonchev Str., Bl. 21, Sofia, 1113, Bulgaria;1. Centro Infant – Pontifícia Universidade Católica do Rio Grande do Sul (PUCRS), Av. Ipiranga, 6690 2° andar, 90610-000 Porto Alegre, RS, Brazil;2. Departamento de Biologia Celular e Molecular, FABIO, Instituto de Pesquisas Biomédicas, PUCRS, Av. Ipiranga, 6690 2° andar, 90610-000 Porto Alegre, RS, Brazil;3. Laboratório de Biologia Molecular, Instituto de Pesquisas Veterinárias Desidério Finamor, Fundação Estadual de Pesquisa Agropecuária, Estrada do Conde, 6000, Eldorado do Sul, RS 92990-000, Brazil;1. School of Pharmacy, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong, China;2. State Key Laboratory of Oncology in South China, Cancer Center, Sun Yat-Sen University, Guangzhou, China;3. School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong, China |
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| Abstract: | Abstract: Four primers for the amplification of mitochondrial DNA of lichen-forming ascomycetes are presented. The primers match the conserved regions U2, U4, and U6, respectively, of mitochondrial small subunit (SSU) ribosomal DNA (rDNA). Polymerase chain reaction using different combinations of the primers produced single amplification products from DNA of eight lichen-forming fungal species but did not amplify DNA of two axenic cultured algal species. The amplification product obtained from Lobaria pulmonaria was sequenced and the 894-bp sequence was compared with the mitochondrial SSU rDNA sequence of Podospora anserina. The two sequences revealed more than 76% identity in the conserved regions U3 to U5 demonstrating that we amplified mitochondrial DNA. The primers matching U2 and U6 yielded amplification products of 800–1000 bp depending on the species examined. The variation observed suggests that mitochondrial SSU rDNA may be useful for phylogenetic analyses of lichen-forming ascomycetes. |
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