scholarly journals Deteksi dan analisis sekuen gen inhibitor proteinase pada beberapa klon kakao harapan tahan penggerek buah kakao dari Sulawesi Selatan Detection and sequence analysis of proteinase inhibitor gene in cacao clones putatively cacao pod borer-tolerant from South Sulawesi

2016 ◽  
Vol 72 (1) ◽  
Author(s):  
Abdul Mollah S. JAYA ◽  
Hajrial ASWIDINNOOR ◽  
Djoko SANTOSO
Keyword(s):  
Sequence Analysis ◽  
Proteinase Inhibitor ◽  
Genomic Dna ◽  
Genetic Similarity ◽  
Sequence Analyses ◽  
South Sulawesi ◽  
Pod Borer ◽  
Special Programs ◽  
Pcr Products ◽  
Alignment Analysis

Summary Cacao is socially and economically an important commodity for Indonesia, in which the cacao plantations have been challenged with a threatening pest, cacao pod borer (CPB). This research aimed to identify and clone PIN (proteinase inhibitor), a gene carrying resistance of plant to some chewing pests like CPB. The methodology included several experiments. Detection of PIN in cacao was done by PCR using PIN-specific heterologous primers and cacao genomic DNA as templates. Cloning vector pGEM-T was utilized to clone the PCR products. Sequence analysis was conducted with BlastX and Blast Special programs from NCBI. Alignment analysis to determine genetic similarity was performed with ClustalW from EBI. Thirteen of the 18 clones tested were detected to have PIN homologs. Two DNA fragments from cacao clones putatively tolerant to CPB, MJ-1 and LW-1, were sequenced. One of them, MJ-1 was cloned. Sequence analyses of the fragments of both cacao clones, indicated that they have PIN homologs and a very closed genetic relation with 96% level of similarity. Ringkasan Kakao adalah komoditas yang secara sosial maupun ekonomi penting bagi Indonesia, dimana perkebunan kakao menghadapi masalah serius hamapenggerek buah kakao (PBK). Penelitian ini bertujuan mengidentifikasi dan mengklon PIN (inhibitor proteinase), gen yang membawa sifat ketahanan tanaman terhadap hama ulat seperti PBK. Metodologinya terdiri dari beberapa percobaan. Deteksi PIN di dalam kakao dikerjakan dengan PCR menggunakan primer heterologous yang spesifik terhadap PIN dan DNA genomik kakao sebagai templetnya. Vektor kloning pGEM-T digunakan untuk mengklon produk PCR. Analisis sekuen dilakukan dengan program BlastX dan Blast spesial dari NCBI. Analisis penjajaran (alignment) untuk menentukan kemiripan genetik menggunakan program ClustalW dari EBI. Tiga belas dari 18 klon kakao yang diuji  menunjukkan adanya  homolog  PIN. Dua DNA fragmen dari klon harapan tahan, MJ-1 dan LW-1, telah ditentukan sekuen nukleotidanya. Satu diantara-nya, MJ-1 berhasil diklon. Analisis sekuen  kedua klon tersebut menunjukkan identitas sebagai homolog PIN dan keduanya memiliki kemiripan genetik yang tinggi.

scholarly journals Deteksi dan analisis sekuen gen inhibitor proteinase pada beberapa klon kakao harapan tahan penggerek buah kakao dari Sulawesi Selatan Detection and sequence analysis of proteinase inhibitor gene in cacao clones putatively cacao pod borer-tolerant from South Sulawesi

2016 ◽  
Vol 72 (1) ◽  
Author(s):  
Abdul Mollah S. JAYA ◽  
Hajrial ASWIDINNOOR ◽  
Djoko SANTOSO
Keyword(s):  
Sequence Analysis ◽  
Proteinase Inhibitor ◽  
Genomic Dna ◽  
Genetic Similarity ◽  
Sequence Analyses ◽  
South Sulawesi ◽  
Pod Borer ◽  
Special Programs ◽  
Pcr Products ◽  
Alignment Analysis

Summary Cacao is socially and economically an important commodity for Indonesia, in which the cacao plantations have been challenged with a threatening pest, cacao pod borer (CPB). This research aimed to identify and clone PIN (proteinase inhibitor), a gene carrying resistance of plant to some chewing pests like CPB. The methodology included several experiments. Detection of PIN in cacao was done by PCR using PIN-specific heterologous primers and cacao genomic DNA as templates. Cloning vector pGEM-T was utilized to clone the PCR products. Sequence analysis was conducted with BlastX and Blast Special programs from NCBI. Alignment analysis to determine genetic similarity was performed with ClustalW from EBI. Thirteen of the 18 clones tested were detected to have PIN homologs. Two DNA fragments from cacao clones putatively tolerant to CPB, MJ-1 and LW-1, were sequenced. One of them, MJ-1 was cloned. Sequence analyses of the fragments of both cacao clones, indicated that they have PIN homologs and a very closed genetic relation with 96% level of similarity. Ringkasan Kakao adalah komoditas yang secara sosial maupun ekonomi penting bagi Indonesia, dimana perkebunan kakao menghadapi masalah serius hamapenggerek buah kakao (PBK). Penelitian ini bertujuan mengidentifikasi dan mengklon PIN (inhibitor proteinase), gen yang membawa sifat ketahanan tanaman terhadap hama ulat seperti PBK. Metodologinya terdiri dari beberapa percobaan. Deteksi PIN di dalam kakao dikerjakan dengan PCR menggunakan primer heterologous yang spesifik terhadap PIN dan DNA genomik kakao sebagai templetnya. Vektor kloning pGEM-T digunakan untuk mengklon produk PCR. Analisis sekuen dilakukan dengan program BlastX dan Blast spesial dari NCBI. Analisis penjajaran (alignment) untuk menentukan kemiripan genetik menggunakan program ClustalW dari EBI. Tiga belas dari 18 klon kakao yang diuji  menunjukkan adanya  homolog  PIN. Dua DNA fragmen dari klon harapan tahan, MJ-1 dan LW-1, telah ditentukan sekuen nukleotidanya. Satu diantara-nya, MJ-1 berhasil diklon. Analisis sekuen  kedua klon tersebut menunjukkan identitas sebagai homolog PIN dan keduanya memiliki kemiripan genetik yang tinggi.

Isolation and sequence analysis of the genomic DNA fragment encoding an aspartic proteinase inhibitor homologue from potato (Solanum tuberosum L.)

1992 ◽  
Vol 20 (2) ◽  
pp. 311-313 ◽  
Author(s):  
Darja Barlič Maganja ◽  
Borut Štrukelj ◽  
Jože Pungerčar ◽  
Franc Gubenšek ◽  
Vito Turk ◽  
...  
Keyword(s):  
Solanum Tuberosum ◽  
Sequence Analysis ◽  
Proteinase Inhibitor ◽  
Genomic Dna ◽  
Solanum Tuberosum L ◽  
Aspartic Proteinase ◽  
Dna Fragment

Mitochondrial Cytochrome b DNA Sequence Variations: An Approach to Fish Species Identification in Processed Fish Products

2005 ◽  
Vol 68 (2) ◽  
pp. 421-425 ◽  
Author(s):  
TIZIANA PEPE ◽  
MICHELE TROTTA ◽  
ISOLINA DI MARCO ◽  
PAOLA CENNAMO ◽  
ANIELLO ANASTASIO ◽  
...  
Keyword(s):  
Sequence Analysis ◽  
Cytochrome B ◽  
Fish Species ◽  
Mitochondrial Cytochrome ◽  
Fish Products ◽  
Community Regulation ◽  
Sequence Variations ◽  
Fish Species Identification ◽  
Mitochondrial Cytochrome B ◽  
Pcr Products

The identification of fish species in food products is problematic because morphological features of the fish are partially or completely lost during processing. It is important to determine fish origin because of the increasing international seafood trade and because European Community Regulation 104/2000 requires that the products be labeled correctly. Sequence analysis of PCR products from a conserved region of the cytochrome b gene was used to identity fish species belonging to the families Gadidae and Merluccidae in 18 different processed fish products. This method allowed the identification of fish species in all samples. Fish in all of the examined products belonged to these two families, with the exception of one sample of smoked baccalà (salt cod), which was not included in the Gadidae cluster.

scholarly journals A new fingerprint method for sequence analysis of chromosomal translocations at the genomic DNA level

Leukemia ◽  
1998 ◽  
Vol 12 (5) ◽  
pp. 758-763 ◽  
Author(s):  
T Leis ◽  
R Repp ◽  
A Borkhardt ◽  
M Metzler ◽  
F Schläger ◽  
...  
Keyword(s):  
Sequence Analysis ◽  
Genomic Dna ◽  
Chromosomal Translocations

scholarly journals Use of short tandem repeat fingerprinting to validate sample origins in hepatitis C virus molecular epidemiology studies

2014 ◽  
Vol 95 (1) ◽  
pp. 66-70 ◽  
Author(s):  
Victoria C. Edwards ◽  
C. Patrick McClure ◽  
Richard J. P. Brown ◽  
Emma Thompson ◽  
William L. Irving ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Sequence Analysis ◽  
Molecular Epidemiology ◽  
Alternative Explanation ◽  
Sequence Analyses ◽  
Distinct Virus ◽  
Virus Sequence ◽  
Epidemiology Studies ◽  
Short Tandem

Sequence analysis is used to define the molecular epidemiology and evolution of the hepatitis C virus. Whilst most studies have shown that individual patients harbour viruses that are derived from a limited number of highly related strains, some recent reports have shown that some patients can be co-infected with very distinct variants whose frequency can fluctuate greatly. Whilst co-infection with highly divergent strains is possible, an alternative explanation is that such data represent contamination or sample mix-up. In this study, we have shown that DNA fingerprinting techniques can accurately assess sample provenance and differentiate between samples that are truly exhibiting mixed infection from those that harbour distinct virus populations due to sample mix-up. We have argued that this approach should be adopted routinely in virus sequence analyses to validate sample provenance.

scholarly journals DNA SEQUENCES OF FRAMESHIFT AND OTHER MUTATIONS INDUCED BY ICR-170 IN YEAST

Genetics ◽  
1985 ◽  
Vol 111 (2) ◽  
pp. 233-241
Author(s):  
Joachim F Ernst ◽  
D Michael Hampsey ◽  
Fred Sherman
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Amino Acid ◽  
Sequence Analysis ◽  
Genetic Analysis ◽  
Dna Sequences ◽  
Induced Mutations ◽  
Sequence Analyses ◽  
Base Pairs ◽  
Yeast Saccharomyces Cerevisiae ◽  
Frameshift Mutations

ABSTRACT ICR-170-induced mutations in the CYC1 gene of the yeast Saccharomyces cerevisiae were investigated by genetic and DNA sequence analyses. Genetic analysis of 33 cyc1 mutations induced by ICR-170 and sequence analysis of eight representatives demonstrated that over one-third were frameshift mutations that occurred at one site corresponding to amino acid positions 29-30, whereas the remaining mutations were distributed more-or-less randomly, and a few of these were not frameshift mutations. The sequence results indicate that ICR-170 primarily induces G·C additions at sites containing monotonous runs of three G·C base pairs. However, some (see PDF) sites within the CYC1 gene were not mutated by ICR-170. Thus, ICR-170 is a relatively specific mutagen that preferentially acts on certain sites with monotonous runs of G·C base pairs.

Polymorphism in the Dirofilaria immitis immunodominant antigen gene

1999 ◽  
Vol 73 (3) ◽  
pp. 265-272 ◽  
Author(s):  
K. Sugane ◽  
K. Nakayama ◽  
H. Kato
Keyword(s):  
Sequence Analysis ◽  
Larval Stage ◽  
Genomic Dna ◽  
Dirofilaria Immitis ◽  
Gene Encoding ◽  
Spliced Leader ◽  
Dna Library ◽  
Immunodominant Antigen ◽  
Genomic Dna Library ◽  
Rapid Amplification

Dg2, a gene encoding a 34 kDa immunodominant antigen of Dirofilaria immitis was cloned and demonstrated to be specifically expressed in the larval stage. In this study, a newly constructed genomic DNA library was screened by hybridization with Dg2. One of the resulting positive clones was similar to Dg2 in the structure of its exonic regions but different in number, position, size and sequence of introns. This was designated DgK. Full-length cDNA was isolated using the rapid amplification of cDNA ends (RACE) method to study the transcript corresponding to DgK. Sequence analysis revealed that the mRNA corresponding to DgK is trans-spliced during post-transcriptional processing because the 5′ end of the amplified cDNA contains seven nucleotides of the nematode-spliced leader (SL) sequence.

Development of an EST-SSR marker in Panax ginseng

2008 ◽  
Vol 5 (2) ◽  
pp. 175-181 ◽  
Author(s):  
Yang Cheng-Jun ◽  
Wang Jun ◽  
Mu Li-Qiang ◽  
Li Shao-Chen ◽  
Liu Guan-Jun ◽  
...  
Keyword(s):  
Panax Ginseng ◽  
Expressed Sequence Tags ◽  
Genomic Dna ◽  
Ssr Marker ◽  
Panax Quinquefolius ◽  
Chain Reaction ◽  
Acanthopanax Senticosus ◽  
Pcr Products ◽  
Polymerase Chain ◽  
Expressed Sequence

AbstractA total of 791 microsatellites (SSRs) were isolated from 7055 Panax ginseng expressed sequence tags (ESTs). According to primer design criteria, 68 primer pairs for EST-SSR were designed. Under an appropriate polymerase chain reaction (PCR) system, all EST-SSR primer pairs were screened against genomic DNA of Ji'anchangbo and Fusong'ermaya from Panax ginseng, and 43 EST-SSR primer pairs out of the above 68 resulted in PCR products. Then, all 43 pairs were detected in nine P. ginseng, two Panax quinquefolius and two Acanthopanax senticosus cultivars for polymorphisms, and 26 pairs (60.47%) were found to be polymorphic, accounting for 38.23% of the total number of designed primer pairs. These results demonstrate the possibility of developing EST-SSR markers using P. ginseng ESTs.

Multiplex Real-Time PCR Detection of Fumonisin-Producing and Trichothecene-Producing Groups of Fusarium Species

2004 ◽  
Vol 67 (3) ◽  
pp. 536-543 ◽  
Author(s):  
B. H. BLUHM ◽  
M. A. COUSIN ◽  
C. P. WOLOSHUK
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Genomic Dna ◽  
Fusarium Species ◽  
Fusarium Spp ◽  
Pcr Assay ◽  
Traditional Methods ◽  
Microbiological Methods ◽  
Pcr Products ◽  
Its Pcr

Some species of Fusarium can produce mycotoxins during food processing procedures that facilitate fungal growth, such as the malting of barley. The objectives of this study were to develop a 5′ fluorogenic (Taqman) real-time PCR assay for group-specific detection of trichothecene- and fumonisin-producing Fusarium spp. and to identify Fusarium graminearum and Fusarium verticillioides in field-collected barley and corn samples. Primers and probes were designed from genes involved in mycotoxin biosynthesis (TRI6 and FUM1), and for a genus-specific internal positive control, primers and a probe were designed from Fusarium rDNA sequences. Real-time PCR conditions were optimized for amplification of the three products in a single reaction format. The specificity of the assay was confirmed by testing 9 Fusarium spp. and 33 non- Fusarium fungal species. With serial dilutions of purified genomic DNA from F. verticillioides, F. graminearum, or both as the template, the detection limit of the assay was 5 pg of genomic DNA per reaction. The three products were detectable over four orders of magnitude of template concentration (5 pg to 5 ng of genomic DNA per reaction); at 50 ng template per reaction, only the TRI6 and FUM1 PCR products were detected. Barley and corn samples were evaluated for the presence of Fusarium spp. with traditional microbiological methods and with the real-time PCR assay. The 20 barley samples and 1 corn sample that contained F. graminearum by traditional methods of analysis tested positive for the TRI6 and internal transcribedspacer (ITS) PCR products. The five corn samples that tested positive for F. verticillioides by traditional methods also were positive for the FUM1 and ITS PCR products. These results indicate that the described multiplex real-time PCR assay provides sensitive and accurate differential detection of fumonisin- and trichothecene-producing groups of Fusarium spp. in complex matrices.

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