eng
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2010-10-01
8
4
213
228
7109
Dopamine-synthesizing neurons: An overview of their development and application for cell therapy
Mossa Gardaneh
mgardaneh@gmail.com
1
Molecular Genetics Group, Stem Cells and Tissue Engineering Committee, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, P.O. Box 14965/161, I.R. Iran.
Cell-gene therapy is a dynamic constituent of novel medical biotechnology. Neurodegenerative disordersin which damage to or demise of specific brain cell types plays central role, are clear examples of diseasecandidate for cell replacement therapy. Dopaminergic (DAergic) neurons biosynthesize dopamine, a vitalneurotransmitter in the central nervous system. Due to the involvement of dopamine in a number of criticalphysiological functions in human and other mammals, disturbed dopamine neurotransmission resulting fromDAergic neuron death or damage causes a few known disorders most prominently Parkinson’s disease (PD).DAergic cell replacement therapies proposed as promising approaches for PD treatment have prompted scientists to thoroughly investigate the embryonic development of DAergic neurons and their function in ordinary life. This review summarizes past and current findings in DAergic neuron development and survival.It also briefly looks at the future prospect of DAergic neuron generation in vitro aiming at clinical applicationsin vivo.
https://www.ijbiotech.com/article_7109_78ea69cbcf942a98946aa028393fdbfc.pdf
Dopamine
dopaminergic
Parkinson disease
cell therapy
eng
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2010-10-01
8
4
229
233
7098
Glucose 6-phosphate dehydrogenase deficiency in Tehran, Zanjan and Sistan-Balouchestan provinces:prevalence and frequency of Mediterranean variant of G6PD
Yousef Mortazavi
1
Fatemeh Mirzamohammadi
2
Majid Teremahi Ardestani
3
Ebrahim Mirimoghadam
4
Tom J Vulliamy
5
Department of Haematology, Zanjan University of Medical Sciences, P.O. Box 4513956111, Zanjan, I.R. Iran.
Student Research Center, Zanjan University of Medical Sciences, P.O. Box 4513956111, Zanjan, I.R. Iran.
Department of Haematology,Yazd University of Medical Sciences, P.O. Box 734, Yazd, I.R. Iran.
Department of Genetics, Zahedan University of Medical School, P.O. Box 98135, Zahedan, I.R. Iran.
Department of Haematology, Division of Investigative Science, Faculty of Medicine, Imperial College of Science, Technology and Medicine, Hammersmith Hospital, London W12 0NN, UK.
Glucose-6-phosphate dehydrogenase (G6PD) deficiency is an enzymopathy affecting about 400 millionpeople worldwide. The distribution of G6PD deficiency and the molecular genetics of this enzyme vary widelyamong different ethnic groups. The aim of this study was to find out the frequency of G6PD deficiency andcharacterize the Mediterranean type mutation in deficient individuals in Turk, Balouch and Fars ethnicgroups in Zanjan, Iranshahr and Tehran provinces. 1500 unrelated male individuals from Zanjan and 305unrelated male students from Iranshahr were screened for G6PD deficiency by fluorescent spot test.Genomic DNA was extracted from deficient individuals and also from 64 G6PD deficient individuals fromTehran city. PCR was used to amplify flanking regions of exons 6 and 7 of this gene. The PCR products weredigested by the MboII enzyme and electrophoresed on 3% agarose gel. Thirty-three (2.2%) individuals wereshown to be deficient for G6PD from Zanjan population. Twenty-four out of 33 (72.8%) of the deficient individuals showed a mutation at nt 563 which is characteristic of Mediterranean type of mutation. Nine individuals were negative for this mutation. Fifty nine (19.3%) individuals of Iranshahr were shown to be deficient for G6PD. At the molecular level, 50 (85%) of the individuals showed Mediterranean type of mutation and 15% were negative for this mutation. Our results from Tehran showed that 47/64 (73.4%) of deficient individuals had Mediterranean type mutation and 26.6% were negative for this mutation. Despite different frequencies exist for deficiency of G6PD in Turk, Balouch and Fars populations, the results of the present study and others have shown that the predominant mutation of G6PD in Iran is of Mediterranean type.
https://www.ijbiotech.com/article_7098_ba67a3881f069b87c88a816ad0b82329.pdf
PCR
G6PD deficiency
Iran
Mediterranean mutation
eng
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2010-10-01
8
4
234
242
7106
Comparison of proliferation and osteoblast differentiation of marrow-derived mesenchymal stem cells on nano- and micro-hydroxyapatite contained composite scaffolds
Mohamadreza Baghaban Eslaminejad
eslami@royaninstitute.org; bagesla@yahoo.com
1
Fatemeh Bagheri
2
Mojgan Zandi
3
Elham Nejati
4
Elham Zomorodian
5
Houri Mivehchi
6
Department of Stem Cell and Developmental Biology, Royan Institute for Stem Cell Biology and Technology, ACECR, P.O. Box 19395-4644, Tehran, I.R. Iran.
Department of Stem Cell and Developmental Biology, Royan Institute for Stem Cell Biology and Technology, ACECR, P.O. Box 19395-4644, Tehran, I.R. Iran.
Department of Polymeric Biomaterials, Iran Polymer and Petrochemical Institute, P.O. Box 115/14965, Tehran, I.R. Iran.
Department of Biomedical Engineering, Amirkabir University of Technology, P.O. Box 15876/4413,Tehran, I.R. Iran.
Department of Stem Cell and Developmental Biology, Royan Institute for Stem Cell Biology and Technology, ACECR, P.O. Box 19395-4644, Tehran, I.R. Iran.
Department of Polymeric Biomaterials, Iran Polymer and Petrochemical Institute, P.O. Box 115/14965, Tehran, I.R. Iran.
Bones constructed by tissue engineering are being considered as valuable materials to be used for regeneration of large defects in natural bone. In an attempt to prepare a new bone construct, in this study, proliferation and bone differentiation of marrow-derived mesenchymal stem cells (MSCs) on our recently developed composite scaffolds of nano-, micro-hydroxyapatite/ poly(l-lactic acid) were compared with purepoly(l-lactic acid) scaffolds. For this purpose, some passaged-3 rat MSCs were three-dimensionally cultivatedon the scaffold surfaces and their morphology was observed with scanning electron microscopy. Cellproliferations on different scaffolds were examined by MTT assays. Osteogenic cultures were establishedwith the scaffolds loaded with MSCs for 21 days at the end of which culture mineralization; the cell alkalinephosphatase (ALP) Level and the relative expression of selected bone specific genes were quantified andcompared to each other. Our results indicated that the cells having been adhered and assumed sphericalmorphology were able to proliferate in all studied scaffolds. The microenvironment provided by nano-scaffoldsappeared much better medium than those of micro-scaffolds and pure PLLA (P < 0.05). The osteogenesisassays indicated to the superiority of nanoscaffolds in supporting MSCs undergoing bone differentiation,which was reflected in high cellular ALP levels, increased bone-related gene expression and enhanced culture mineralization. Collectively, the bone construct prepared with nano-hydroxyapatite/ poly (llactic acid) scaffold and proliferated MSCs would be suitable candidate for use in bone regenerative medicine.
https://www.ijbiotech.com/article_7106_f8c55b4546361489a38c51aab3ff29e4.pdf
Nano- and micro-hydroxyapatite
poly (llactic acid)
Mesenchymal stem cell
Bone differentiation
Cell proliferation
Scaffold
eng
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2010-10-01
8
4
243
251
7110
Inhibition of ackA and pta genes using two specific antisense RNAs reduced acetate accumulation in batch fermentation of E. coli BL21 (DE3)
Nahid Bakhtiari
1
Manouchehr Mirshahi
2
Valiollah Babaeipour
3
Nader Maghsoudi
4
Biochemistry Group, Faculty of Biological Sciences, Tarbiat Modares University, P.O. Box 1411-5111, Tehran, I.R. Iran.
Biochemistry Group, Faculty of Biological Sciences, Tarbiat Modares University, P.O. Box 1411-5111, Tehran, I.R. Iran.
Biochemical Engineering Group, Biotechnology Research Center, P.O. Box 15857-1774, Tehran, I.R.Iran.
Neuroscience Research Center, Shahid Beheshti University of Medical Science, P.O. Box 19615-1178, Tehran, I.R. Iran.
Expression of foreign proteins in E. coli is normally inhibited by exogenous production of acetate. To overcomethis problem, various strategies have been proposed and tested to reduce the extent of acetate accumulation.Although these strategies can improve the outcome, the implementation of their proposed techniquesis not practical. Because to achieve optimal results, it requires extremely tight control conditionsand the actual cost is very high. Furthermore, a simple knockout mutation of the target metabolic pathwaywould not be appropriate because the acetate pathway plays an important physiological role in E. coli. In thisstudy, we employed an antisense RNA strategy as an elaborated metabolic engineering tool to partially blockbiosynthesis of two major acetate pathway enzymes, acetate kinase (ACK) and phosphotransacetylase(PTA). The fragments of antisense cassette were cloned sequentially in pBluescriptsk+ and completedcassette subcloned in pLT10T3. The function of this cassette was evaluated with RT-PCR and ACK andPTA assay. The effect of cassette on cell physiology was monitored by determination of optical density, glucose consumption and acetate production. We found that the antisense method partially reduced mRNA levels of the target genes, lowered the concentration of acetate in culture media and increased growth rateand final cell density in antisense-regulated strain. This strategy could provide us with a useful, inexpensiveand practical tool to achieve a large-scale protein production system.
https://www.ijbiotech.com/article_7110_9766287b03132b355e7c34e82695bdef.pdf
Foreign protein
Antisense RNA technology
Acetate pathway
ACK
PTA
eng
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2010-10-01
8
4
252
262
7099
Serratia marcescens B4A chitinase product optimization using Taguchi approach
Mandana Zarei
zarei.mandana@yahoo.com
1
Saeed Aminzadeh
minzade@nigeb.ac.ir
2
Hossein Zolgharnein
zolgharnine@yahoo.com;zolgharnein@kmsu.ac.ir
3
Alireza Safahieh
safahieh@hotmail.com
4
Ahmad Ghoroghi
ghoroghiahmad@yahoo.com
5
Abbasali Motallebi
6
Morteza Daliri
7
Abbas Sahebghadam Lotfi
lotfi_ab@modares.ac.ir
8
Department of Animal and Marine Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), P.O. Box 14965/161, Tehran, I.R. Iran.
Department of Animal and Marine Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), P.O. Box 14965/161, Tehran, I.R. Iran.
Faculty of Marine Sciences, Khorramshahr Marine Sciences and Technology University, P.O. Box 669, Khorramshahr, I.R. Iran.
Faculty of Marine Sciences, Khorramshahr Marine Sciences and Technology University, P.O. Box 669, Khorramshahr, I.R. Iran.
Iranian Fisheries Research Organization, P.O. Box 14155/6116, Tehran, I.R. Iran.
Iranian Fisheries Research Organization, P.O. Box 14155/6116, Tehran, I.R. Iran.
Department of Animal and Marine Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), P.O. Box 14965/161, Tehran, I.R. Iran.
Department of Animal and Marine Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), P.O. Box 14965/161, Tehran, I.R. Iran.
Chitinase production by newly isolated Serratia marcescens B4A was optimized following Taguchi’sarray methods. Twenty-three bacterial isolates were screened from shrimp culture ponds in the South ofIran. A chitinase-producing bacterium was isolated based on it’s ability to utilize chitin as the sole carbonsource. The isolate designated as B4A, was identified as Serratia marcescens based on its 16S rRNAsequence and key morphological, physiological and biochemical characteristics. The cultivation of Serratiamarcescens B4A in the appropriate liquid medium resulted in production of high levels of chitinase. Themalt extract and colloidal chitin represented the best nitrogen and carbon sources, respectively. Chitinaseproduction by Serratia marcescens B4A was optimized following the Taguchi orthogonal array (OA) for thedesign of experiments (DOE). Statistical experimental design via the Taguchi method was applied to determine the optimal levels of physical parameters and key media components in the medium, such as temperature, pH, NaCl and chitin concentrations. The results of this study showed that temperature of 30ºC, pH 7.9, NaCl 0.1% (w/v) and chitin 1% (w/v) are optimal conditions for this protocol.
https://www.ijbiotech.com/article_7099_229a09fc874016b1155794ffc98e3bb4.pdf
Chitinase
isolation
Screening
Serratia marcescens B4A
Taguchi method
eng
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2010-10-01
8
4
263
269
7113
Callus induction and shoot regeneration from epicotyl explants of ethnomedicinally important Caesalpinia bonduc (L.) Roxb
Meena Kochappan Cheruvathur
1
John Britto
2
Thuruthiyil Dennis Thomas
3
Post graduate and Research Department of Botany, St. Thomas College, Pala, Arunapuram (P.O), PIN- 686 574, Kottayam (Dt.), Kerala, India.
The Rapinat Herbarium and Centre for Molecular Systematics and Taxonomy, St. Joseph’s College (Autonomous). Tiruchirappalli-620 002, Tamil Nadu, India.
Post graduate and Research Department of Botany, St. Thomas College, Pala, Arunapuram (P.O), PIN- 686 574, Kottayam (Dt.), Kerala, India
An efficient plant regeneration method was established for Caesalpinia bonduc (L.) Roxb. by culturingimmature epicotyl explants. Morphogenic calli were initiated from 96% of epicotyl explants on MS mediumsupplemented with BA (6-benzylaminopurine) 4.0 mg/l and NAA (Naphthalene acetic acid) 1.0 mg/l. The calliformed were excised and subcultured on MS medium fortified with 1 mg/l 2, 4-dichlorophenoxyacetic acid (2,4-D) for further proliferation. Maximum percent organogenesis (84%) and average shoots per culture(5.6) was observed on MS medium fortified with 3.0 mg/l BA and 1.0 mg/l IAA (indole-3- acetic acid).Addition of indole-3-butyric acid (IBA) in ½ MS medium favored rooting of recovered shoots. Out of 30 rootedshoots transferred to soil 27 survived after acclimatization.
https://www.ijbiotech.com/article_7113_00b7943ff95e26416774571d983314c2.pdf
Caesalpinia bonduc
Callus
Epicotyl
Ethnomedicinal plant
Regeneration
eng
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2010-10-01
8
4
270
274
7111
Bacterial overexpression of the human interleukin-2 in insoluble form via the pET Trx fusion system
Sohrab Boozarpour
1
Majid Sadeghizadeh
sadeghma@modares.ac.ir
2
Mohammad Ali Shokrgozar
mashokrgozar@pasteur.ac.ir
3
Saman Hosseinkhani
saman_h@modares.ac.ir
4
Seyed Abbas Shojaosadati
shoja@modares.ac.ir
5
Sara Gharavi
6
Ghasem Ahangari
gh.ahangari@yahoo.com
7
Bijan Ranjbar
8
Department of Genetics, Faculty of Biological Sciences, Tarbiat Modares University, P.O. Box 14115-154, Tehran, I.R. Iran.
Department of Genetics, Faculty of Biological Sciences, Tarbiat Modares University, P.O. Box 14115-154, Tehran, I.R. Iran.
National Cell Bank of Iran, Pasteur Institute, P.O. Box 1316943551, Tehran, I.R. Iran.
Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, P.O. Box 14115- 154,Tehran, I.R. Iran.
Biotechnology Group of Chemical Engineering, Faculty of Engineering, Tarbiat Modares University, P.O. Box 14115-143, Tehran, I.R. Iran.
Department of Biology, Alzahra University, P.O. Box 1993891176, Tehran, I.R. Iran.
Department of Medical Genetics, National Institute of Genetic Engineering and Biotechnology, P.O. Box 14155-6343, Tehran, I.R. Iran.
Department of Biophysics, Faculty of Biological Sciences, Tarbiat Modares University, P.O. Box 14115-154, Tehran, I.R. Iran.
Selection of a system for successful recombinant protein production is important. The aim of this study wasto produce high levels of human interleukin-2 (hIL-2) in soluble form. To this end, the pET32a vector inEscherichia coli BL21 (DE3) was used as an expression system, since it was previously used for the productionof mouse IL-2 in soluble form. The results indicated that contrary to expectations, the expressed proteinwas in the form of inclusion bodies and perhaps amino acid differences between human and mouse IL-2 should be determinant. The hIL-2 protein is a small peptide, therefore its recovery as a biologically functional protein by the process of refolding may be feasible and could lead to high yields at the industrial scale.
https://www.ijbiotech.com/article_7111_7967f761c8f5942c6967289c5fb76165.pdf
Human IL-2
Soluble protein
Inclusion body
Overexpression
E. coli
eng
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2010-10-01
8
4
275
279
7112
Functional screening of phosphatase-encoding genes from bacterial sources
Mohammad Reza Sarikhani
1
Mohammad Ali Malboobi
malboobi@nigeb.ac.ir
2
Nasser Aliasgharzad
3
Ralf Greine
4
Bagher Yakhchali
bahar@nigeb.ac.ir
5
National Institute of Genetic Engineering and Biotechnology, P.O. Box 14965/161, Tehran, I.R. Iran.
National Institute of Genetic Engineering and Biotechnology, P.O. Box 14965/161, Tehran, I.R. Iran.
University of Tabriz, Faculty of Agriculture, Department of Soil Science, P.O. Box 5166616471, Tabriz, I.R. Iran.
Federal Research Institute of Nutrition and Food, Haid-und-Neu-Straße 9, 76131 Karlsruhe, Germany.
National Institute of Genetic Engineering and Biotechnology, P.O. Box 14965/161, Tehran, I.R. Iran.
Phosphatase (APase) enzymes including phytases have broad applications in diagnostic kits, poultryfeeds, biofertilizers and plant nutrition. Because of high levels of sequence diversity among phosphatases,an efficient functional screening method is a crucial requirement for the isolation of the encodinggenes. This study reports a functional cloning screening method for the isolation of APase-encoding genesfrom bacterial genomic libraries in a medium containing a chromogenic substrate. The method was optimizedto distinguish the desired signal from the background chromosomal APase activity. This screening method led to the isolation of two novel APase-encoding genes from Pseudomonas putida with no similarities to the known genes in the databases, indicating successful implementation of the developed method.
https://www.ijbiotech.com/article_7112_9f7825604d12b9afcad2bc63307c4101.pdf
Pseudomonas putida
Phosphatase
Phytase
Phytate
BCIP