Papers by Xavier DANIEL
Agrobacterium tumefaciens telah digunakan secara luas pada penelitian molekuler tanaman dan pemul... more Agrobacterium tumefaciens telah digunakan secara luas pada penelitian molekuler tanaman dan pemuliaan tanaman sejak tahun 1983 untuk kepentingan rekayasa genetika. Umumnya A.tumefaciens diinfeksi pada diskus daun sedangkan menginjeksi A.tumefaciens ke jaringan daun dapat dijadikan alternatif, metode ini disebut Agroinfiltrasi. Agroinfiltrasi lebih cepat dan mudah dibandingkan teknis diskus daun karena dapat mengukur ekspresi transgene sejak 2-4 hari setelah infiltrasi. Sedangkan metode diskus daun memerlukan waktu 2-4 bulan untuk analisa ekspresi transgene. Pada penelitian ini, Agroinfiltrasi daun dilakukan pada Sindoro salah satu jenis tembakau Temanggung. Variabel agroinfiltrasi adalah densitas inokulum A.tumefaciens C58C1(pMp90, pEGAD) dan waktu ko-kultivasi setelah infiltrasi. Kuantifikasi Green Fluorescent Protein (GFP) ekstrak daun Sindoro dibandingkan kontrol negatif dapat mencapai 123% pada OD600=0.06 saat 24 jam setelah infiltrasi. Sebagian agroinfiltrasi daun Sindoro juga diregenerasi pada media Murashige dan Skoog (MS) mengandung BASTA 5 ppm sebagai agen seleksi. Tunas transforman muncul pada hari ke-47 setelah inokulasi di media MS seleksi transforman.
KnE Life Sciences, Sep 11, 2017
The xynB gene of Bacillus subtilis subsp. spizizenii W23 is predicted to encode a xylan 1,4-beta-... more The xynB gene of Bacillus subtilis subsp. spizizenii W23 is predicted to encode a xylan 1,4-beta-xylosidase. Application of XynB enzymes in industries is wide. Production of this enzyme in its host cells is naturally restricted by repression process. It will give certain beneficial to over-expressed the enzymes in other host-cells under inducing promoter. This study aimed to clone the xynB gene from Bacillus subtilis subsp. spizizenii W23, to pMMB67EH plasmid, and to over-express the xynB gene in Escherichia coli Origami as host cells. The xynB gene was successfully amplified by polymerase chain reaction (PCR) technique using a pair of primers flanking the gene sequence and chromosomal DNA of the W23 strain as a template. The xynB gene inserted in recombinant plasmid was confirmed by PCR detection using primers pair's specific for xynB gene and for the vector, then continued by restriction analyses. The result showed that transformants clone 9 and 10 bear the recombinant pMMB-xynB plasmid. The xylanase activity of xynB gene in Escherichia coli Origami clone 10 was detected by sodium-dodecyl-sulfate polyacrylamide gel analyses and with addition of isopropyl-D-thio-galactoside (IPTG) as an inducer. The protein seem to be overexpressed as intra-and extra-cellular protein detected on SDS-PAGE gel. Result from xylan degrading activity on Luria-Bertani-xylan-IPTG plate with addition of Congo Red, showed that the cells with pMMB-xynB recombinant plasmid have clear zone around the colonies while the transformant bearing an empty plasmid showed no clear zone. It could be concluded that the xynB gene of Bacillus subtilis subsp.spizizenii W23 has been successfully been cloned on pMMB67EH plasmid and over-expressed in the Escherichia coli Origami cells as intra-and extra-cellular protein, as observed on SDS-PAGE gel analysis. The protein has activity on xylan degradation.
Cantaloupe Melon (Cucumis melo L.) is a fruit plant riching in antioxidant beta-carotene. Perhaps... more Cantaloupe Melon (Cucumis melo L.) is a fruit plant riching in antioxidant beta-carotene. Perhaps, production of beta-carotene can also be achieved by first initiating callus culture of the plant in suitable medium, propagating them, and extracting the compound it produce. This research was a preliminary effort aimed to examine it. The results of this research showed that the optimum medium for callus culture initiation from cotyledon of Cantaloupe Melon and its propagation was Murashige Skoog (MS) with the addition of 1 mg/L benzyl adenine (BA) and 1.5 mg/L napthalene acetic Acid (NAA). The beginning of stationary phase on calluses' growth curve was chosen as a harvest time of calluses, based on the theory that beta-carotene as a secondary metabolite is usually be produced much at that phase. The results demonstrated that stationary phase began at the end of week fourth, so the time was decided as the harvest time of calluses. Beta-carotene was then extracted from calluses by maceration technique. The existence of the compound in extract was tested using Thin Layer Chromatography (TLC) and Fourier Transformed-Infrared Spectroscopy (FTIR). The data showed that the compound existed in the extract. The concentration of the compound in it was needed to determine further.
Springer eBooks, 2017
Syzygium cumini (Linn.) Skeels, known as jamblang, belongs to the family of guava (Myrtaceae). Th... more Syzygium cumini (Linn.) Skeels, known as jamblang, belongs to the family of guava (Myrtaceae). The plant can be found in India, Southeast Asia and Eastern Africa. This plant has a potential use for treatment of type 2 diabetes mellitus. Although still visible, especially in rural areas, it is becoming very rare in big cities. Nowadays, not many people use the fruit of this plant because of its scarcity. To overcome the problem, in this research, jamblang plant was propagated in vitro using the leaves as explants to produce callus cultures instead of the regeneration of whole plants. The sterilization was conducted by immersing them in the Mushi Guard solution (5 mL/L), followed by the immersion in the solution of NaOCl. The callus cultures were here induced using WPM (Woody Plant Medium) with different concentrations of growth hormones, i.e., naphthalene acetic acid (NAA) and benzylaminopurine (BAP), in order to determine their optimum concentrations for the induction. The results showed the optimum media of calluses induction was WPM with NAA 5 ppm without any addition of BAP with an induction time of 30 days and percentage of callus cultures induction up to 90%. The optimum media for calluses growth was also WPM with the addition of NAA 5 ppm. Their average growth index was 2.212 after the 30 days of subculturing.
Mata kuliah Teknik Analisa DNA mengajarkan kepada mahasiswa mengenai teknik-teknik analisa Biolog... more Mata kuliah Teknik Analisa DNA mengajarkan kepada mahasiswa mengenai teknik-teknik analisa Biologi Molekuler, khususnya DNA. Namun, rancangan perangkat kurikulum dan metode pengajaran yang sebelumnya diterapkan masih menitikberatkan pada prosedur, sehingga minat dan motivasi belajar mahasiswa menjadi rendah. Inovasi pembelajaran ini bertujuan untuk menata ulang perangkat kurikulum berdasarkan metode pembelajaran kontekstual serta merancang metode dan media pembelajaran yang mengacu pada Student Centered Learning (SCL) yang diharapkan dapat meningkatkan minat dan motivasi belajar mahasiswa. Pembelajaran kontekstual dilakukan melalui pemberian case study dan proyek yang mengaplikasikan teknik analisa DNA untuk mencari solusi atas suatu permasalahan. Hasil implementasi inovasi pembelajaran ini menunjukkan rerata capaian ketuntasan belajar sebesar 61,28% dan peningkatan motivasi belajar mandiri mahasiswa sebesar 51,25%. Selain itu, juga terjadi peningkatan kepuasan mahasiswa terhadap kinerja dosen. Output inovasi pembelajaran ini juga dihasilkan perangkat kurikulum yang baru dan beberapa media pembelajaran seperti software offline pembelajaran, handout kuliah dan buku ajar. Kata kunci: inovasi pembelajaran, pembelajaran kontekstual, student centered learning PENDAHULUAN Mata kuliah Teknik Analisa DNA mengajarkan kepada mahasiswa mengenai teknik-teknik analisa DNA yang mendukung beberapa mata kuliah lain seperti Genetika Molekuler, Rekayasa Genetika dan Skripsi. Mata kuliah ini akan diikuti oleh mata kuliah Praktikum Teknik Analisa DNA yang dilaksanakan pada semester berikutnya, sehingga selain memahami teori dari teknik yang digunakan, nantinya diharapkan mahasiswa juga mahir menerapkannya di laboratorium. Mata kuliah ini ditujukan bagi mahasiswa semester 4 meskipun tidak menutup kemungkinan brought to you by CORE View metadata, citation and similar papers at core.ac.uk
This study aimed to clone thexynB gene from Bacillussubtilissubsp. spizizeniiW23,encodinga xylan ... more This study aimed to clone thexynB gene from Bacillussubtilissubsp. spizizeniiW23,encodinga xylan l,4-beta-xylosidase to pMMB67EH plasmid which then be used to transformed EscherichiacoliDH-5a and Origami host cells. The xynBgenewas amplifiedby polymerasechain reaction (PCR) technique using a pair of primers flanking the gene sequence,and chromosomal DNA of the W23 strain as a template. Analyses of the recombinant plasmid were done by restriction analyses, and PCR detection. The result showed that the xynB has been cloned on pMMB67EH vector. The recombinant plasmid contained the xynB gene which was confirmed by restriction analyses and by PCR detection using primers pair's specific for the xynB gene and for the vector. The xylanaseactivity of xynB gene in E.coliDH-5a and Origami host cells was assayedon luria-Bertani-xylan plate qualitatively with addition of isopropyl B-D-thio-galactoside (IPTG) as an inducer.Uponsprayingwith Congored, the cells bearing the pMMB-xynB recombinant plasmid showed a xylan-degrading activity by the appearance ofclear zanearound the colonies whilethe transformant bearing an empty plasmid showed no clear zone. It could be concluded that the cloning Arocess was succeeded.
Mata kuliah Teknik Analisa DNA mengajarkan kepada mahasiswa mengenai teknik-teknik analisa Biolog... more Mata kuliah Teknik Analisa DNA mengajarkan kepada mahasiswa mengenai teknik-teknik analisa Biologi Molekuler, khususnya DNA. Namun, rancangan perangkat kurikulum dan metode pengajaran yang sebelumnya diterapkan masih menitikberatkan pada prosedur, sehingga minat dan motivasi belajar mahasiswa menjadi rendah. Inovasi pembelajaran ini bertujuan untuk menata ulang perangkat kurikulum berdasarkan metode pembelajaran kontekstual serta merancang metode dan media pembelajaran yang mengacu pada Student Centered Learning (SCL) yang diharapkan dapat meningkatkan minat dan motivasi belajar mahasiswa. Pembelajaran kontekstual dilakukan melalui pemberian case study dan proyek yang mengaplikasikan teknik analisa DNA untuk mencari solusi atas suatu permasalahan. Hasil implementasi inovasi pembelajaran ini menunjukkan rerata capaian ketuntasan belajar sebesar 61,28% dan peningkatan motivasi belajar mandiri mahasiswa sebesar 51,25%. Selain itu, juga terjadi peningkatan kepuasan mahasiswa terhadap ki...
La reponse hypersensible ou hr, caracterisee par la mort des cellules vegetales au point d'in... more La reponse hypersensible ou hr, caracterisee par la mort des cellules vegetales au point d'infection, et conduisant au confinement de l'agent pathogene, est generalement associee a la resistance des plantes aux agents pathogenes. Cette reponse fait appel a un programme genetique de mort cellulaire, et si les evenements precoces conduisant au declenchement de la hr sont de mieux en mieux connus, l'execution du programme de mort cellulaire demeure en revanche une enigme. Le travail presente dans ce manuscrit est focalise sur un acteur potentiel de ce programme chez arabidopsis thaliana, le gene atmyb30. Ce gene avait ete isole au laboratoire sur la base de son expression, induite specifiquement lors d'une interaction incompatible avec la bacterie pathogene xanthomonas campestris pv. Campestris, et appartient a la famille des facteurs transcriptionnels de type myb. Une etude precise de son expression a montre qu'elle est specifique, precoce et transitoire lors de la...
Proceedings of the National Academy of Sciences, 2010
A novel myb oncogene homologue (AtMYB30) has been isolated by differential screening of a cDNA li... more A novel myb oncogene homologue (AtMYB30) has been isolated by differential screening of a cDNA library prepared from Xanthomonas campestris pv. campestris (X. campestris)-inoculated Arabidopsis thaliana cells cultured in the presence of cycloheximide. AtMYB30 is a single-copy gene, and the encoded protein contains a MYB domain highly homologous to other plant and animal MYB proteins. Analyses of transcript levels in A. thaliana plants, or in cultured A. thaliana cells infected with either virulent or avirulent strains of the pathogens X. campestris and Pseudomonas syringae pv. tomato, showed that maximal levels of transcription of this gene occurred during the hypersensitive response. Furthermore, in A. thaliana mutants affected in the control of cell death initiation (lsd3, lsd4 and lsd5), constitutive expression or expression in lesion-positive plants was observed, while in suppressors of the mutations lsd5 and lsd4, AtMYB30 transcripts did not accumulate. However, AtMYB30 expression could not be detected in the lsd1 mutant, which was hyperresponsive to cell death initiators and unable to limit the extent of cell death, whatever the environmental conditions. The results presented here suggest a strong correlation between AtMYB30 and genetically controlled cell death, with a role in the initiation of cell death rather than in the limitation of its extent. Our results further indicate that the lsd mutants constitute an appropriate genetic model for studying the role of this gene in hypersensitive cell death, and their relation to different steps of the pathway(s) leading to cell death.
Plant …, Jan 1, 2012
An attack of plants by pathogens or treatment with certain resistance-inducing compounds can lead... more An attack of plants by pathogens or treatment with certain resistance-inducing compounds can lead to the establishment of a unique primed state of defense. Primed plants show enhanced defense reactions upon further challenge with biotic or abiotic stress. Here, we report that the primed state in Arabidopsis (Arabidopsis thaliana) is still functional in the next generation without additional treatment. We compared the reactions of Arabidopsis plants that had been either primed with β-amino-butyric acid (BABA) or with an avirulent isolate of the bacteria Pseudomonas syringae pv tomato (PstavrRpt2). The descendants of primed plants showed a faster and higher accumulation of transcripts of defense-related genes in the salicylic acid signaling pathway and enhanced disease resistance upon challenge inoculation with a virulent isolate of P. syringae. In addition, the progeny of primed plants was also more resistant against the oomycete pathogen Hyaloperonospora arabidopsidis. When transgenerationally primed plants were subjected to an additional priming treatment, their descendants displayed an even stronger primed phenotype, suggesting that plants can inherit a sensitization for the priming phenomenon. Interestingly, this primed to be primed phenotype was much reduced in the Arabidopsis β-amino-butyric acid priming mutant ibs1 (induced BABA sterility1). Our results demonstrate that the primed state of plants is transferred to their progeny and confers improved protection from pathogen attack as compared to the descendants of unprimed plants.
Proceedings of the …, Jan 1, 2002
Hypersensitive response (HR) is a programmed cell death that is commonly associated with disease ... more Hypersensitive response (HR) is a programmed cell death that is commonly associated with disease resistance in plants. Among the different HR-related early induced genes, the AtMYB30 gene is specifically, rapidly, and transiently expressed during incompatible interactions between Arabidopsis and bacterial pathogens. Its expression was also shown to be deregulated in Arabidopsis mutants affected in the control of cell death initiation. Here, we demonstrate that overexpression in Arabidopsis and tobacco of
Proceedings of the …, Jan 1, 2004
The circadian clock controls numerous physiological and molecular processes in organisms ranging ... more The circadian clock controls numerous physiological and molecular processes in organisms ranging from fungi to human. In plants, these processes include leaf movement, stomata opening, and expression of a large number of genes. At the core of the circadian clock, the central oscillator consists of a negative autoregulatory feedback loop that is coordinated with the daily environmental changes, and that generates the circadian rhythms of the overt processes. Phosphorylation of some of the central oscillator proteins is necessary for the generation of normal circadian rhythms of Drosophila, humans, and Neurospora, where CK1 and CK2 are emerging as the main protein kinases involved in the phosphorylation of PER and FRQ. We have previously shown that in Arabidopsis, the protein kinase CK2 can phosphorylate the clock-associated protein CIRCADIAN CLOCK ASSOCIATED 1 (CCA1) in vitro. The overexpression of one of its regulatory subunits, CKB3, affects the regulation of circadian rhythms. Whether the effects of CK2 on the clock were due to its phosphorylation of a clock component had yet to be proven. By examining the effects of constitutively expressing a mutant form of the Arabidopsis clock protein CCA1 that cannot be phosphorylated by CK2, we demonstrate here that CCA1 phosphorylation by CK2 is important for the normal functioning of the central oscillator.
Teaching Documents by Xavier DANIEL
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Papers by Xavier DANIEL
Teaching Documents by Xavier DANIEL