scholarly journals Penyisipan Gen Inhibitor α-amilase pada Plasmid Biner pCambia 1301

2016 ◽  
Vol 1 (2) ◽  
pp. 45
Author(s):  
Edy Listanto ◽  
Sutrisno Sutrisno ◽  
Saptowo J. Pardal ◽  
M. Herman
Keyword(s):  
Molecular Biology ◽  
Research And Development ◽  
Genetic Resources ◽  
Restriction Enzymes ◽  
Restriction Pattern ◽  
E Coli ◽  
Colony Number ◽  
The Right ◽  
Biology Laboratory ◽  
Binary Plasmid

<p class="p1">The experiment was conducted at the Molecular Biology Laboratory of the Indonesian Center for Agricultural Biotechnology and Genetic Resources Research and Development, Bogor. The objective was to construct <em>-ai </em>gene on a binary plasmid <em>p</em>Cambia 1301. This experiment was carried out using construction method by ligation process between fragments of <em>α-ai </em>gene from <em>p</em>TA<span class="s1">3 </span>plasmid and <em>p</em>Cambia 1301 on <em>Hind</em>III site. The result of ligant transformation into <em>E. coli </em>DH5<em>α </em>was 182 surviving colonies on YEP medium containing kanamycin. DNA samples were obtained from 60 randomly selected colonies. The restriction pattern was tested by digesting each DNA sample using <em>Hind</em>III showed colonies containing two fragments expected of sizes wich are 11.837 and 4.887 kb. Two colonies are predicted containing of <em>α-ai </em>gene on its the binary plasmid. Advanced tests using restriction enzymes <em>Bam</em>HI and <em>Xba</em>I showed two directions (right and left) of <em>α-ai </em>gene. The right direction was shown by <em>p</em>Cambia-<em>α-ai</em>1 from colony number 43. This plasmid showed expected fragments of sizes 13.485 and 3.219 kb when digested with <em>Bam</em>HI and two fragments of sizes 15.421 and 1.303 kb when digested with <em>Xba</em>I. The left direction was shown <em>p</em>Cambia-<em>α-ai</em>2 from colony number 58. This plasmid also demon-strated expected fragments of sizes 15.026 and 1.698 kb when digested with <em>Bam</em>HI and two fragments of sizes 13.082 and 3.642 kb when digested with <em>Xba</em>I. Both <em>p</em>Cambia-<em>α-ai</em>1 and <em>p</em>Cambia-<em>α-ai</em>2 were transformed into <em>A. tumefaciens </em>LBA4404.</p>

The Central Importance of E. coli and λ‎ Phage in the New Molecular Biology

2021 ◽  
pp. 130-135
Author(s):  
Thomas E. Schindler
Keyword(s):  
Molecular Biology ◽  
Genetic Recombination ◽  
Model Organism ◽  
Restriction Enzymes ◽  
Model Organisms ◽  
Host Restriction ◽  
E Coli ◽  
Jumping Genes ◽  
K 12 ◽  
Convenient Model

This chapter considers two of the most important legacies of the Lederbergs’ pioneering work: the discoveries of the model organisms that would dominate molecular biology, E. coli and λ‎ bacteriophage. The Lederbergs’ introduction of E. coli as a convenient model organism shifted the direction of molecular genetics. Barbara McClintock’s discovery of jumping genes remained unappreciated for decades, until the field of molecular biology caught up to validate her transposable elements in bacteria. The discovery of restriction enzymes—the molecular scissors for precisely cutting DNA at specific sites, a prerequisite for genetic recombination techniques—emphasized the versatility of bacteriophage λ‎ as a powerful experimental tool. The discovery of specialized transduction by Larry Morse and Esther Lederberg hinted at the mechanisms of “host restriction.” Werner Arber and Daisy Dussoix discovered restriction endonucleases by building upon Esther Lederberg’s research with λ‎ phage and the differences between E. coli B and K-12.

Pengembangan buku ajar praktikum mata kuliah morfologi tumbuhan di laboratorium biologi

2016 ◽  
Vol 1 (1) ◽  
pp. 1
Author(s):  
Suraida Suraida
Keyword(s):  
Research And Development ◽  
Learning Process ◽  
Teaching And Learning ◽  
Lesson Plan ◽  
Plant Morphology ◽  
The State ◽  
State Institute ◽  
Plan Execution ◽  
Islamic Studies ◽  
Biology Laboratory

Abstrak Penelitian ini dilakukan karena proses pembelajaran di laboratorium Biologi IAIN STS Jambi yang masih minim sarana prasarana yang ada di laboratorium, sehingga menghambat proses pembelajaran khususnya untuk mata kuliah Morfologi Tumbuhan. Penelitian ini bertujuan mengembangkan buku ajar praktikum dan mengetahui praktikalitasnya. Jenis Penelitian ini adalah penelitian pengembangan (Research and Development) dengan menggunakan 4-D Models yang terdiri dari 4 tahap yaitu Define, Design, Develop dan Disseminate. Karena adanya keterbatasan waktu dan biaya maka tahap disseminate tidak dilakukan. Produk yang dikembangkan berupa buku ajar praktikum yang divalidasi oleh validator. Produk yang telah divalidasi dan dinyatakan valid oleh validator, kemudian diujicobakan pada proses pembelajaran yang bertujuan untuk melihat nilai praktis buku ajar praktikum di laboratorium Biologi. Analisis data yang digunakan adalah data deskriptif untuk memvalidasi perangkat pembelajaran oleh pakar pendidikan. Selain itu juga diteliti data praktikalitas penggunaan perangkat pembelajaran ini yang diperoleh dari observasi dosen dan respon siswa. Nilai validitas produk 83,31% yang dikategorikan valid. Sementara nilai kepraktisan berdasarkan data observasi keterlaksanaan SAP, angket respon dosen dan siswa dikategorikan sangat baik atau sangat praktis. Penelitian menyimpulkan bahwa perangkat pembelajaran di Laboratorium Biologi yang dikembangkan adalah valid dan sangat praktis digunakan baik dosen maupun siswa. Kata Kunci : Pengembangan, buku ajar praktikum, laboratorium biologi Abstract [The development of a course book for plant morphology at biology laboratory] This research was triggered by the limited facilities of the biology laboratory at the State Institute of Islamic Studies Sulthan Thaha Saifuddin Jambi which became a constrain in the teaching and learning process of Plant Morphology classroom sessions. The objective of this research was to develop a course book as well as to reveal its practicality. The researcher did a research and development using 4-D Models consisting of four stages namely; define, design, develop, and disseminate. Considering the limitation of time and finance, the disseminate stage was not executed. The test revealed the validity score of the product was 83,31% which categorized as good. For its practicality, the product was considered as very good based on observation of lesson plan execution and lecturers’ and students’ response. In summary, the course book developed for the course at Biology Laboratory was categorized as valid and practical to be used by both students and lecturers. Keywords: development, a course book, biology laboratory

Viability of Edwardsiella tarda and Esherichia coli preserved with glycerol-tryptone soy broth (TSB) kept at freezing temperature

2013 ◽  
Vol 1 (2) ◽  
pp. 154
Author(s):  
Abdur Rohman ◽  
Frans Ijong ◽  
I K Suwetja
Keyword(s):  
Research And Development ◽  
Germ Plasm ◽  
Freezing Temperature ◽  
Experimental Methods ◽  
Edwardsiella Tarda ◽  
Diagnostic Tools ◽  
Glycerol Concentration ◽  
E Coli ◽  
Esherichia Coli ◽  
Escherchia Coli

Preservation of bacteria carried out in relation to the collection and preservation of germ plasm microbe is useful for research and development or for the establishment of diagnostic tools. Glycerol is a good preservation media but it is not known what doses should be used for effective preservation.  This research used two experimental  methods consisting of 2 factors and 3 treatments. This study aimed to find the best glycerol concentration that can be used to preserve Edwarsiella tarda and Escherchia coli in the -20ºC environment, to understand the viability of bacteria after being preserved and to describe the characteristics of the preserved bacteria. Treatments applied were 10%, 15% and 20%  glycerol in TSB. Viability of the bacteria was analyzed after 7, 14, 28, 35, and 42 days of preservation. Results showed that E.coli bacteria preserved in 15%  glycerol had the highest viability, i.e. 84% and preserved in 10% glycerol had the lowest viability, i.e. 80%. But for E. tarda bacteria preserved in 10% glycerol had the highest viability, i.e. 1.83% and preseved in 15% glycerol had the lowest viability, i.e. 0,55%. Preservasi bakteri dilakukan dalam kaitannya dengan koleksi dan konservasi plasma nutfah mikroba yang berguna untuk penelitian dan pengembangan atau untuk pembentukan alat diagnosa. Gliserol merupakan bahan preservasi yang baik, tetapi belum diketahui dosis yang baik dan efektif untuk preservasi bakteri Edwarsiella tarda dan Escherchia coli pada suhu -20ºC. Penelitian ini dilakukan dengan metode eksperimen yang terdiri dari 2 faktor dan 3 taraf perlakuan, masing-masing perlakuan dengan 3 kali ulangan, media preservasi yang digunakan adalah TSB dan gliserol dengan konsentrasi 10%, 15% dan 20%. Parameter yang diukur adalah viabilitas dan kecocokan/penyimpangan karakteristik biokimia. Penelitian ini dilaksanakan di Laboratorium Balai Karantina Ikan Pengendalian Mutu dan Keamanan Hasil Perikanan Manado, dari bulan September sampai dengan November 2013. Tujuan Penelitian ini adalah untuk menentukan konsentrasi gliserol dalam TSB sebagai media preservasi yang efektif dan efisien pada bakteri  Edwarsiella tarda dan Escherchia coli yang dipreservasi dengan suhu -20ºC dan disimpan selama 42  hari. Hasil penelitian menunjukkan adanya penurunan laju pertumbuhan bakteri selama preservasi. Persentase viabilitas  bakteri E. coli yang tertinggi selama preservasi diperoleh dengan penggunaan gliserol konsentrasi 15% dengan jumlah 84% dan yang terendah adalah dengan penggunaan konsentrasi 10% yakni sebesar 80%, sedangkan untuk E. tarda persentase viabilitas  bakteri yang tertinggi selama preservasi diperoleh dengan penggunaan gliserol konsentrasi 10% dengan jumlah 1,83% dan yang terendah adalah dengan penggunaan konsentrasi 15% yakni sebesar 0,55%. Berdasarkan uji statistik analisis variasi (ANAVA) didapat hasil F hitung E. tarda dan E. coli yang lebih besar  dari FTabel dengan tingkat kepercayaan 95 %.

scholarly journals Lowering DNA binding affinity of SssI DNA methyltransferase does not enhance the specificity of targeted DNA methylation in E. coli

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Krystyna Ślaska-Kiss ◽  
Nikolett Zsibrita ◽  
Mihály Koncz ◽  
Pál Albert ◽  
Ákos Csábrádi ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Dna Binding ◽  
Binding Affinity ◽  
Dna Methyltransferase ◽  
Restriction Enzymes ◽  
Dna Binding Protein ◽  
E Coli ◽  
Dna Binding Affinity ◽  
Higher Eukaryotes

AbstractTargeted DNA methylation is a technique that aims to methylate cytosines in selected genomic loci. In the most widely used approach a CG-specific DNA methyltransferase (MTase) is fused to a sequence specific DNA binding protein, which binds in the vicinity of the targeted CG site(s). Although the technique has high potential for studying the role of DNA methylation in higher eukaryotes, its usefulness is hampered by insufficient methylation specificity. One of the approaches proposed to suppress methylation at unwanted sites is to use MTase variants with reduced DNA binding affinity. In this work we investigated how methylation specificity of chimeric MTases containing variants of the CG-specific prokaryotic MTase M.SssI fused to zinc finger or dCas9 targeting domains is influenced by mutations affecting catalytic activity and/or DNA binding affinity of the MTase domain. Specificity of targeted DNA methylation was assayed in E. coli harboring a plasmid with the target site. Digestions of the isolated plasmids with methylation sensitive restriction enzymes revealed that specificity of targeted DNA methylation was dependent on the activity but not on the DNA binding affinity of the MTase. These results have implications for the design of strategies of targeted DNA methylation.

scholarly journals PROSPEK PENERAPAN PP NO. 35 TAHUN 2007 SEBAGAI PENGATURAN INSENTIF PERPAJAKAN UNTUK MENDORONG PENINGKATAN R&D

2009 ◽  
Vol 39 (2) ◽  
pp. 207
Author(s):  
Brian Amy Prastyo
Keyword(s):  
Research And Development ◽  
Government Policy ◽  
Government Regulation ◽  
Domestic Market ◽  
Huge Amount ◽  
Tax Incentive ◽  
The Republic ◽  
Conducting Research ◽  
The Right ◽  
Recent Government

AbstrakThis article elaborates on tax incentive for research and development area.In recent government policy of the Republic Indonesia has governed underthe Government Regulation number 35 year 2007. The author deems thatthat regulation has some fundamentals weakness. The policy toward taxincentive commonly should make corporation can save in huge amount. Thataspect is also contained disadvantage to be abused. Hence abuse throughduty free privilege could happen like on imported goods selling which isimported by that privilege abuse in domestic market. Under the author sightsthen government ought to launch any exclusive policy for certain companythat has significant program to conducting research and development. Thenthe author advises to protect that policy also must be secured by worthysystem of incentive's application will be approved to the right one only

scholarly journals Malakoplakia of the prostate masquerading as locally advanced prostate cancer on mpMRI

2015 ◽  
Vol 9 (11-12) ◽  
pp. 910 ◽  
Author(s):  
Robert Thomas Dale ◽  
Michael Metcalfe ◽  
Silvia Chang ◽  
Edward Jones ◽  
Peter Black
Keyword(s):  
Locally Advanced ◽  
Specific Antigen ◽  
Transrectal Ultrasound ◽  
Locally Advanced Prostate Cancer ◽  
E Coli ◽  
Intravenous Antibiotics ◽  
Left Apex ◽  
Left Base ◽  
The Right ◽  
Extraprostatic Extension

A 66-year-old man was referred for urological evaluation for an abnormal digital rectal exam (cT2a, subtle nodule at left base, 121 cc prostate) and an elevated prostate specific antigen (PSA) of 8.0 ng/ml. Subsequent 12-core transrectal ultrasound (TRUS)- guided biopsy revealed Gleason 3+4 adenocarcinoma in seven of 12 cores, including all six cores on the right side and one core at the left apex. No extraprostatic extension was identified. Postbiopsy, the patient developed urinary retention requiring a catheter, as well as an Escherichia coli (E. coli) urinary tract infection (UTI) requiring hospitalization and intravenous antibiotics.

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