Isolasi fragmen gen LIPASE dari kapang Absidia corymbifera, Rhizopus oryzae dan Rhizopus oligosporus Isolation of LIPASE gene fragment from Absidia corymbifera, Rhizopus oryzae and Rhizopus oligosporus fungi

AbstractDiversification of oil palm products, suchas healthy oil, needs lipase sustainability as abiocatalist. Many attempts have beendeveloped to produce lipase, includingintensive exploration and screening of severalspecies of molds. Genetic engineering for overexpression of LIPASE gene in the selectedmold is considered to be the potentialapproach for efficient production of thisenzyme. This research was aimed to isolate theLIPASE gene fragment of Indonesianindigenous fungi, namely Absidia corymbifera,Rhizopus oryzae and R. oligosporus by meansof RT-PCR (Reverse Transcriptase PolymeraseChain Reaction) technique using heterologousprimers. The result showed that a cDNAfragment of 462 bp has been amplified andisolated from the three fungi with differentconcentration. The highest quantity was foundfrom A. corymbifera. The RT-PCR productsisolated from A. corymbifera was cloned,sequenced and analyzed for its homology to thesequence of LIPASE gene from other species.BLAST analysis showed that the DNA sequenceof the cloned RT-PCR product derived fromA. corymbifera was highly homologous withLIPASE gene from Rhizopus niveus.AbstraksDiversifikasi produk kelapa sawit, sepertiminyak sehat (healthy oil) memerlukanketersediaan lipase sebagai biokatalis. Berbagaiupaya untuk produksi lipase telah dikembang-kan, termasuk eksplorasi dan skrining terhadapbeberapa spesies kapang secara intensif.Rekayasa genetika untuk mengoverekspresi-kan gen LIPASE pada kapang hasil skriningtersebut dipandang merupakan satu pendekatanpotensial untuk produksi enzim ini secaraefisien. Penelitian ini bertujuan untukmengisolasi fragmen gen LIPASE dari tigakapang indigenous Indonesia, yaituA. corymbifera, R. oryzae dan R. oligosporus,menggunakan teknik RT-PCR (ReverseTranscriptase Polymerase Chain Reaction).Hasil penelitian menunjukkan bahwa fragmencDNA sepanjang 462 bp dari ketiga kapangtelah diisolasi, masing-masing dengankuantitas yang berbeda. Hasil tertinggidiperoleh dari kapang A. corymbifera. ProdukRT-PCR dari A. corymbifera diklon, disekuenkemudian dianalisis homologinya dengansekuen gen LIPASE dari spesies lain. AnalisisBLAST menunjukkan bahwa sekuen DNA dariproduk RT-PCR terklon yang berasal dariA. corymbifera memiliki homologi tinggidengan gen LIPASE dari Rhizopus niveus.

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RT-PCR amplification of a Rhizopus oryzae lactate dehydrogenase gene fragment

Enzyme and Microbial Technology ◽

10.1016/s0141-0229(00)00319-7 ◽

2001 ◽

Vol 28 (2-3) ◽

pp. 259-264 ◽

Cited By ~ 9

Author(s):

Erdogan E. Hakki ◽

Mahinur S. Akkaya

Keyword(s):

Lactate Dehydrogenase ◽

Rhizopus Oryzae ◽

Pcr Amplification ◽

Gene Fragment ◽

Rt Pcr ◽

Dehydrogenase Gene

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An outbreak of viral gastroenteritis associated with consumption of sandwiches: implications for the control of transmission by food handlers

Epidemiology and Infection ◽

10.1017/s0950268898001150 ◽

1998 ◽

Vol 121 (3) ◽

pp. 615-621 ◽

Cited By ~ 107

Author(s):

U. D. PARASHAR ◽

L. DOW ◽

R. L. FANKHAUSER ◽

C. D. HUMPHREY ◽

J. MILLER ◽

...

Keyword(s):

Food Handler ◽

Common Source ◽

Viral Gastroenteritis ◽

Food Handlers ◽

Rt Pcr ◽

Source Of Infection ◽

Food Borne ◽

Pcr Product ◽

Stool Specimens ◽

Polymerase Chain

Although food handlers are often implicated as the source of infection in outbreaks of food-borne viral gastroenteritis, little is known about the timing of infectivity in relation to illness. We investigated a gastroenteritis outbreak among employees of a manufacturing company and found an association (RR=14·1, 95% CI=2·0–97·3) between disease and eating sandwiches prepared by 6 food handlers, 1 of whom reported gastroenteritis which had subsided 4 days earlier. Norwalk-like viruses were detected by electron microscopy or reverse transcriptase-polymerase chain reaction (RT-PCR) in stool specimens from several company employees, the sick food handler whose specimen was obtained 10 days after resolution of illness, and an asymptomatic food handler. All RT-PCR product sequences were identical, suggesting a common source of infection. These data support observations from recent volunteer studies that current recommendations to exclude food handlers from work for 48–72 h after recovery from illness may not always prevent transmission of Norwalk-like viruses because virus can be shed up to 10 days after illness or while exhibiting no symptoms.

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Application of RT-PCR for Indexing Avocado Sunblotch Viroid

Plant Disease ◽

10.1094/pdis.1997.81.9.1023 ◽

1997 ◽

Vol 81 (9) ◽

pp. 1023-1026 ◽

Cited By ~ 21

Author(s):

R. J. Schnell ◽

D. N. Kuhn ◽

C. M. Ronning ◽

D. Harkins

Keyword(s):

Rt Pcr ◽

Amplification Product ◽

Chain Reaction ◽

Pcr Assay ◽

Specific Primers ◽

Pcr Product ◽

Polymerase Chain ◽

Dideoxynucleotide Chain Termination Method ◽

Slab Gels ◽

Avocado Sunblotch Viroid

A method for the routine detection of avocado sunblotch viroid (ASBVd) in nucleic acid extracts of infected avocado tissues by reverse transcription-polymerase chain reaction (RT-PCR) was developed using ASBVd-specific primers. Amplified cDNA products were analyzed by electrophoresis on nondenaturing 6% polyacrylamide slab gels. The size of the major RT-PCR product from ASBVd-infected tissue was estimated to be 250 bp. This product was absent from amplified extracts of uninfected tissue. The amplification product from ASBVd was sequenced by the dideoxynucleotide chain termination method, and the sequence was over 97% identical to the published sequence. The RT-PCR assay is sensitive enough to allow viroid detection without requiring large amounts of tissue, highly purified ASBVd, or molecular hybridization.

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First Report of the Green Rosette Variant of Groundnut Rosette Disease in Kenya

Plant Disease ◽

10.1094/pdis.1999.83.8.782a ◽

1999 ◽

Vol 83 (8) ◽

pp. 782-782 ◽

Cited By ~ 2

Author(s):

A. W. Wangai ◽

S. S. Pappu ◽

H. R. Pappu ◽

N. Okoko ◽

C. M. Deom ◽

...

Keyword(s):

Sub Saharan Africa ◽

Satellite Rna ◽

Rt Pcr ◽

Crop Season ◽

Production Constraints ◽

Pcr Product ◽

Arachis Hypogaea L ◽

Disease Surveys ◽

Polymerase Chain ◽

Sub Saharan

Groundnut (Arachis hypogaea L.) is an important food crop in sub-Saharan Africa. One of the major production constraints is groundnut rosette disease, which is caused by a complex of two viruses, groundnut rosette assistor luteovirus (GRAV) and groundnut rosette umbravirus (GRV) together with the associated satellite RNA (satRNA) (1). Two main forms of the disease have been described: chlorotic and the green rosette. Variants of the satRNA have been shown to be largely responsible for the different forms of the disease (1). Chlorotic rosette has been the predominant form in all of sub-Saharan Africa while green rosette has been reported in the western and southern regions of Africa (2). During the 1997-1998 crop season, disease surveys conducted in Kenya showed the incidence of the rosette disease in farmers' fields to be 24 to 40% in a total of 23 fields surveyed in the western regions of the country (Homabay, Kendubay, Kisumu) and 30% in 8 fields sampled in the Rift Valley (Cheplamus, Marigat) regions. Representative peanut plants showing rosette symptoms were analyzed for the presence of GRV by reverse transcription polymerase chain reaction (RT-PCR). With primers specific to a portion of ORF4 of GRV RNA (3), RT-PCR gave a product of expected size (approximately 300 bp). The PCR product was cloned in pGEM-T vector and sequenced. The sequenced region showed 89% nucleotide sequence identity with published GRV sequences. Green rosette was observed on groundnut cultivars Nyaela Red and Homabay Local in the Kendu Bay region. The incidence of the green rosette was 5.3% of the plants with rosette symptoms. References: (1) A. F. Murant and I. K. Kumar. Ann. Appl. Biol. 117:85, 1990. (2) R. A. Naidu et al. Ann. Appl. Biol. 132:525, 1998. (3). M. E. Taliansky et al. J. Gen. Virol. 77:2335, 1996.

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Simple Quantitative Reverse Transcribed-Polymerase Chain Reaction (RT-PCR) Method Involving Recombinant RNA Generated by a False-Priming PCR Product

The Journal of Biochemistry ◽

10.1093/oxfordjournals.jbchem.a124662 ◽

1994 ◽

Vol 116 (6) ◽

pp. 1198-1201 ◽

Cited By ~ 4

Author(s):

Makoto Sato ◽

Mutsuhiko Mizobuchi ◽

Koji Murao ◽

Miho Tamaki ◽

Jiro Takahara

Keyword(s):

Polymerase Chain Reaction ◽

Rt Pcr ◽

Chain Reaction ◽

Pcr Product ◽

Pcr Method ◽

Polymerase Chain

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Cloning, expression and bioactivity of chicken receptor activator of NF-κB ligand (chRANKL)

Chinese Journal of Agricultural Biotechnology ◽

10.1017/s1479236208002131 ◽

2008 ◽

Vol 5 (3) ◽

pp. 205-209

Author(s):

Wang Yan ◽

Hou Jia-Fa

Keyword(s):

Primary Cultures ◽

Rt Pcr ◽

Gene Splicing ◽

His Tag ◽

Pcr Product ◽

Overlap Extension ◽

Receptor Activator ◽

Relative Molecular Weight ◽

Polymerase Chain ◽

Sequence Encoding

AbstractUsing reverse transcription polymerase chain reaction (RT-PCR) and gene splicing by overlap extension (SOE-PCR), the DNA sequence encoding the chicken receptor activator of NF-κB ligand (RANKL) was amplified, cloned into the expressing plasmid pET-32a(+) with His-tag, and highly expressed in Escherichia coli. The purified protein was then added to primary cultures of chicken osteoclasts to observe its bioactivity. The results showed that the size of the PCR product was 1200 bp, which was consistent with the expected size, and the relative molecular weight of the induced protein was 64 kDa. Western blotting indicated that induced protein could react with anti-His antibody. It was also found that the induced protein can stimulate mature chicken osteoclasts to resorb bone.

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Detection of Enteroviral RNA Sequences in Different Water Samples

Water Science & Technology ◽

10.2166/wst.1993.0349 ◽

1993 ◽

Vol 27 (3-4) ◽

pp. 219-222 ◽

Cited By ~ 6

Author(s):

D. Tougianidou ◽

K. Botzenhart

Keyword(s):

Polymerase Chain Reaction ◽

Reverse Transcription ◽

Water Samples ◽

Test System ◽

Rt Pcr ◽

Chain Reaction ◽

Rna Sequences ◽

Pcr Product ◽

Filter Unit ◽

Polymerase Chain

Viruses were isolated from different water samples by Sterivex-Filtration. The nucleic acids were isolated in the filter unit and purified by phenol-chloroform extraction and ethanol precipitation. Reverse transcription and polymerase chain reaction (RT/PCR) were performed with primer pairs complementary to sequences of the enteroviral 5’ noncoding region. Amplified sequences were detected by hybridisation with an oligonucleotide complementary to a part of the PCR product. The test system seems to be sensitive and specific In the detection of enteroviral RNA.

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Karakterisasi gen penyandi lipase dari kapang Rhizopus oryzae dan Absidia corymbifera Characterization of gene encoding lipase from fungus Rhizopus oryzae and Absidia corymbifera

E-Journal Menara Perkebunan ◽

10.22302/iribb.jur.mp.v74i1.118 ◽

2016 ◽

Vol 74 (1) ◽

Author(s):

Riza A PUTRANTO ◽

Djoko SANTOSO ◽

. TRI-PANJI ◽

. SUHARYANTO ◽

Asmini BUDIANI

Keyword(s):

Genomic Dna ◽

Rhizopus Oryzae ◽

High Homology ◽

Lipase Gene ◽

Gene Encoding ◽

Absidia Corymbifera ◽

Blast Analysis ◽

Genomic Dnas ◽

Fat Hydrolysis

SummaryLipase is a group of enzymes which catalyze fat hydrolysis. Lipase is recently used to produce diacylglycerol (DAG) from triacylglycerol (TAG). Lipase can be used to produce healthy oil. Having a rich biodiversity, Indonesia has the opportunity to produce lipase using indigenous microbes, such as molds. This research aimed to detect LIPASE gene on several strains of molds employing PCR technique. Genomic DNAs were isolated from four strains of molds (M. sitophila, R. oryzae, R. microsporus, and A. corymbifera). Heterologous primers for LIPASE were designed based on the conserved region of 12 LIPASE sequences accessed from GenBank and used to amplify the genomic DNA resulted in a 466 bp fragmen. BLAST analysis showed that the bands of DNAs have high homology with common lipase protein in several strains of Rhizopus.Ringkasan Lipase merupakan kelompok enzim yang berfungsi sebagai biokatalis hidrolisis lemak. Lipase banyak digunakan untuk konversi triasilgliserol (TAG) menjadi diasilgliserol (DAG). Penggunaan lipase penting untuk produksi minyak sehat (healthy oil). Indonesia dengan keanekaragaman hayati tinggi berpeluang besar mengembangkan produksi lipase dari mikroba lokal, salah satunya adalah kapang. Deteksi gen merupakan langkah awal dalam upaya peningkatan produksi lipase melalui rekayasa genetika. DNA genomik empat galur kapang (M. sitophila, R. oryzae, R. microsporus, dan A. corymbifera) telah berhasil diisolasi. Sepasang primer heterologous telah berhasil dirancang berdasarkan daerah terkonservasi 12 sekuen gen LIPASE dari GenBank. Amplikon DNA yang diperoleh pada PCR menggunakan pasangan primer RLP memiliki panjang 466 bp. Analisis BLAST memperlihatkan bahwa amplikon PCR memiliki homologi yang tinggi dengan protein LIPASE beberapa galur Rhizopus.

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A case report of rhino-facial mucormycosis in a non-diabetic patient with COVID-19: a systematic review of literature and current update

BMC Infectious Diseases ◽

10.1186/s12879-021-06625-3 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Faezeh Mohammadi ◽

Milad Badri ◽

Shapoor Safari ◽

Nima Hemmat

Keyword(s):

Rhizopus Oryzae ◽

Fungal Infections ◽

Lung Involvement ◽

Review Of Literature ◽

Rt Pcr ◽

Predisposing Factor ◽

Case Presentation ◽

Pcr Product ◽

Wide Range ◽

Nasal Biopsy

Abstract Background COVID-19 disease may be associated with a wide range of bacterial and fungal infections. We report a patient with COVID-19 infection who developed rhino-facial mucormycosis during treatment with corticosteroids. Case presentation A 59-year-old non-diabetic male patient was admitted with a diagnosis of COVID-19 based on positive RT-PCR and CT of the lungs. Due to sever lung involvement, he was treated with methylprednisolone. The patient was re-admitted to hospital, due to nasal obstruction and left side facial and orbital swelling, several days after discharge. In sinus endoscopic surgery, debridement was performed and the specimens were sent to pathology and mycology laboratories. A nasal biopsy showed wide hyphae without septa. The sequenced PCR product revealed Rhizopus oryzae. Despite all medical and surgical treatment, the patient died. In addition, the characteristics of patients with COVID-19-associated mucormycosis were reviewed in 44 available literatures. In most studies, diabetes mellitus was the most common predisposing factor for mucormycosis. Conclusion Our report highlights the need for assessing the presence of mucormycosis in patients with COVID-19 and also it shows that physicians should consider the potential for secondary invasive fungal infections in COVID-19 cases.

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