Genomic analysis of a virulent and a less virulent strain of the entomopathogenic fungus Beauveria bassiana, using restriction fragment length polymorphisms

1991 ◽  
Vol 37 (7) ◽  
pp. 534-541 ◽  
Author(s):  
John M. Kosir ◽  
James M. MacPherson ◽  
George G. Khachatourians
Keyword(s):  
Beauveria Bassiana ◽  
Restriction Fragment ◽  
Fragment Length ◽  
Entomopathogenic Fungus ◽  
Genomic Dna ◽  
Tubulin Gene ◽  
Virulent Mutant ◽  
Fungus Beauveria Bassiana ◽  
Restriction Fragment Length ◽  
Β Tubulin

The genomic DNA of two strains of the entomopathogenic fungus Beauveria bassiana, strain GK2016, a "wild type" (virulent), and strain GK2051, a less virulent mutant to grasshoppers, was digested with 12 restriction endonucleases. Gel electrophoresis conditions were established to show restriction fragment length patterns visually in the digested DNA stained with ethidium bromide. The less virulent mutant was generated by ultraviolet illumination of conidiospores at a 95% lethal dose. Both strains of the fungi were identical in morphology as well as in 16 of 22 API-ZYM kit enzyme assays. Differences in levels of total enzyme activity were observed for esterase, esterase–lipase, (β-galactosidase, chitinase, and protease. A Neurospora crassa β-tubulin gene (heterologous gene) and two homologous DNA probes (pJK16 and pJK18) hybridized to several specific DNA bands in B. bassiana strain GK2016 but not in strain GK2051. Strain GK2051 gave different restriction fragment length patterns when compared with its parent strain. Taken together, the data show restriction fragment length differences between the genomic DNA of the two strains, including the loss of some DNA sequences from the mutant strain, which may be involved in pathogenicity. Finally, B. bassiana GK2016 contains a β-tubulin gene with at least partial homology to that of N. crassa. Key words: restriction fragment length polymorphism, entomopathogen, Beauveria bassiana, bioinsecticide, filamentous fungi.

Restriction fragment length polymorphisms of Anisakinae larvae

1989 ◽  
Vol 63 (4) ◽  
pp. 269-274 ◽  
Author(s):  
Kazuo Sugane ◽  
Liu Qing ◽  
Tadashi Matsuura
Keyword(s):  
Restriction Fragment ◽  
Fragment Length ◽  
Genomic Dna ◽  
Paratenic Host ◽  
Restriction Fragment Length Polymorphisms ◽  
Type I ◽  
Type Ii ◽  
Length Polymorphisms ◽  
Paratenic Hosts ◽  
Restriction Fragment Length

ABSTRACTThe analysis of restriction fragment length polymorphisms (RFLPs) was applied to distinguish several kinds of Anisakinae larvae, Anisakis larvae (type I) collected from two different paratenic hosts, Anisakis larvae (type II) and Contracaecum larvae. The patterns of the two different paratenic host-derived DNA of Anisakis larva (I) were exactly the same in hybridized fragments generated by six endonucleases. The quite different patterns in RFLPs of genomic DNA were observed among the Anisakis larva (I), Anisakis larva (II) and Contracaecum larvae. The results suggest that the RFLPs analysis may be useful for distinguishing Anisakinae larvae and clarifying the relationships between Anisakis larvae and their adult worms.

scholarly journals IDENTIFICATION OF ROSE CULTIVARS BY RESTRICTION FRAGMENT LENGTH POLYMORPHISMS

HortScience ◽  
1991 ◽  
Vol 26 (5) ◽  
pp. 484b-484
Author(s):  
Sriyani Rajapakse ◽  
Mark Hubbard ◽  
Albert Abbott ◽  
Robert Ballard ◽  
John Kelly
Keyword(s):  
Restriction Fragment ◽  
Fragment Length ◽  
Genomic Dna ◽  
Cultivar Identification ◽  
Restriction Fragment Length Polymorphisms ◽  
Repeated Sequences ◽  
Dna Library ◽  
Dichotomous Key ◽  
Length Polymorphisms ◽  
Restriction Fragment Length

Restriction Fragment Length Polymorphisms (RFLPs) were investigated in rose cultivars as a means of reliable cultivar identification. A random genomic DNA library was generated by shotgun cloning HindIII digested fragments of DNA extracted from rose cultivar Confection into pUC8 plasmid of Escherichia coli strain JM 83. Compared to genomic clones carrying low or highly repeated sequences, clones with moderately repeated sequences were most effective in cultivar identification. These clones were identified by hybridizing rose DNA fragments from the library with genomic DNA from `Confection'. Clones with moderately repeated copy sequences were used as probes to detect the presence of RFLPs by Southern hybridization of EcoRI digested genomic DNA of various rose cultivars. Several of these probes have revealed RFLPs useful in cultivar identification. By using a combination of two or more of these probes most of the rose cultivars compared at this time can be identified. A dichotomous key useful in identification of rose cultivars was prepared from RFLPs displayed by 3A9 probe.

scholarly journals Optimasi Isolasi Genom untuk Analisis Keragaman Mikrob pada Fermentasi Singkong "Peyem" dengan Teknik Terminal Restriction Fragment Length Polymorphism (T-RFLP)

2013 ◽  
Vol 18 (1) ◽  
Author(s):  
Tati Barus
Keyword(s):  
Restriction Fragment Length Polymorphism ◽  
16S Rdna ◽  
Restriction Fragment ◽  
Length Polymorphism ◽  
Fragment Length ◽  
Genomic Dna ◽  
Fragment Length Polymorphism ◽  
Dna Purification ◽  
Restriction Fragment Length

"Peyem" merupakan salah satu pangan fermentasi Indonesia. Kualitas pangan fermentasi bergantung pada aktivitas mikrob yang terdapat selama proses fermentasi berlangsung. Salah satu teknik molekuler yang telah banyak digunakan untuk menganalisis komunitas mikrob pada suatu habitat adalah teknik Terminal–Restriction Fragment Lenght Polymorphism (T-RFLP). Metode isolasi genom dan jenis primer yang digunakan pada saat PCR penting pada teknik T- RFLP dalam mengkaji komunitas mikrob. Oleh sebab itu, penelitian ini bertujuan untuk membandingkan empat metode isolasi genom dan membandingkan penggunaan dua set primer dalam mengkaji komunitas bakteri dari "Peyem" dengan teknik T-RFLP. Genom komunitas bakteri diisolasi dengan menggunakan empat metode, yaitu: 1) QIAamp DNA Stool Mini Kit (G1), 2) QIAamp DNA Stool Mini Kit + lisozim (G2), 3) Genomic DNA Purification Kit (G3), dan 4) Genomic DNA Purification Kit + lisozim (G4). Untuk mengamplifikasi 16S rDNA digunakan dua set primer, yaitu: 1) primer 27F-FAM dan 1492R, 2) primer 63F-FAM dan 1387R. Hasil penelitian menunjukkan isolasi genom dengan metode G4 menghasilkan konsentrasi genom tertinggi (330,20 ng/µl) dibandingkan metode G1, G2, dan G3 (163,50 ng/µl; 183,25 ng/µl, dan 260,80 ng/µl). Primer 27F-FAM menghasilkan jumlah peak yang lebih tertinggi (264) dibandingkan dengan primer 63F-FAM (177). Jumlah peak TRF pada teknik TRFLP menggambarkan keragaman komunitas mikrob. Dengan demikian isolasi genom dengan Genomic DNA Purification Kit + lysozyme dan penggunaan pasangan primer 27F-FAM-1492R adalah yang terbaik untuk menganalisis komunitas bakteri dari "Peyem" dengan teknik T-RFLP.Kata kunci: Genom, Primer, T-RFLP, Mikrob, "Peyem"

Use of genomic DNA restriction fragment length differences to identify nematode species

Parasitology ◽  
1985 ◽  
Vol 90 (1) ◽  
pp. 137-144 ◽  
Author(s):  
J. Curran ◽  
D. L. Baillie ◽  
J. M. Webster
Keyword(s):  
Restriction Fragment ◽  
Fragment Length ◽  
Genomic Dna ◽  
Nematode Species ◽  
Agarose Gel Electrophoresis ◽  
Closely Related Species ◽  
Size Dependent ◽  
Dna Restriction ◽  
Endonuclease Digestion ◽  
Restriction Fragment Length

Restriction endonuclease digestion of genomic DNA generates DNA fragments of unique size, dependent upon the particular base sequence. Following fractionation by agarose gel electrophoresis, repetitive DNA can be visualized as distinct bands in stained gels and the restriction fragment length of such bands used as diagnostic characters. Restriction fragment length differences were detected between species within the genera Trichinella, Caenorhabditis, Romanomermis, Steinernema (syn. Neoaplectana) and Meloidogyne. This technique provides a new tool for the taxonomist, which is independent of phenotypic variation and it enables the rapid and reliable separation of closely related species.

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