eng
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2004-04-01
2
2
75
83
6935
Endophytic Fungi, Characteristics and Their Potential for Genetic Manipulation
Aghafakhr Mirlohi
1
Mohammad Reza Sabzalian
2
Mojtaba Khayyam Nekouei
3
Department of Plant Breeding, Isfahan University of Technology, Isfahan, Iran.
Department of Plant Breeding, Isfahan University of Technology, Isfahan, Iran.
Agriculture Biotechnology Institute of Central Region of I.R. Iran.
Unique benign endophytes from Ascomycets have wide distribution among grass species. The symbioticfungi enhance plant characters including performance, insect and mammalian deterrence, nematode resistance and tolerance to drought, salt and other biotic and abiotic stresses. Endophytes from genusNeotyphodium (Acremonium) are of the major focus than their ancestors, and Epichloe species, becausethe formers have lost their sexual reproduction. Therefore they should be genetically stable, and mostimportantly, they cannot disassociate from host tissues, and are transferred vertically. They are maternallyinherited and are therefore attractive for genetic transformation without the concern about geneescape. Some marker genes have been successfully transferred to endophyte Neotyphodium coenophialumand Neotyphodium lolii existing in Festuca arundinacea Schreb. and Lolium perenne L., respectively.Furthermore, gene silencing has been proved to be feasible for eliminating traits, which are economicallyharmful. Methods of direct DNA uptake using polyethylene glycol (PEG) and electroporation have beenfound to be useful in transformation of these fungi. Transgenic fungi can be reinserted into the host withoutneed to tissue culture. The endophytic genes responsible for a specific trait can be isolated and transferred to grass species or other microorganisms for direct exploitation of secondary metabolites and endophytic enzymes. Considering advancements in this filed, endophytes can open new horizons faced to scientists and biotechnologist to use them as a surrogate target of transformation.
https://www.ijbiotech.com/article_6935_724bf7d6beddf379db85387c7ad1e249.pdf
Endophyte
Molecular marker
Neotyphodium
Transformation
eng
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2004-04-01
2
2
84
89
6928
Cloning and Expression of the Coat Protein Gene of Barley Yellow Dwarf Virus-PAV in Escherichia coli
Masoud Shams-bakhsh
shamsbakhsh@modares.ac.ir
1
Robert Henry Symons
2
Department of Plant Pathology, Tarbiat Modarres University, P.O. Box: 14115-143, Tehran, I.R. Iran.
Department of Plant Science, Waite Institute, University of Adelaide, Glen Osmond, SA 5064, Australia.
Due to the restriction of Barley yellow dwarf virus (BYDV)-PAV particles to the phloem tissue and very low virus titers, purification of the virus is difficult. The aim of this study was to prepare antibody against viralcoat protein without purifying the virus. To produce recombinant coat protein, the coding sequence wasfirst amplified from a PAV full-length cDNA clone by polymerase chain reaction (PCR), ligated into a vector(pBluescript SK+) to check the sequence, and subcloned into an expression vector (pGEX-2T). It wasthen transformed into Escherichia coli DH5α by electroporation. The open reading frame 3 (ORF3) waslinked in-frame to the gene encoding glutathione-Stransferase (GST; 26 kDa) and expression induced byIPTG. The expressed coat protein was purified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) for use as an immunogen. The antisera to BYDV-PAV recombinant coat protein reacted inWestern blot analysis with partially purified BYDV-PAV. These antisera were also used to detect BYDV-PAV byimmunogold electron microscopy of thin section of barley tissues. The results indicated that BYDV-PAV coatprotein can be produced in high yields by E. coli, which provides the ability of simple purification, and becauseof proper antigencity, can be exploited for diagnostic applications.
https://www.ijbiotech.com/article_6928_29ead82d79f50238f1f3877b5df856e4.pdf
Barley yellow dwarf virus
Expression vector
polyclonal antibodies
E. coli
Coat protein
PAV
eng
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2004-04-01
2
2
90
96
6929
The Effect of Trap Plants on the Population Diversity of Bradyrhizobium japonicum
Amir Lakzian
alakzian@yahoo.com
1
Eden Bromfield
2
Soil Science Department. Agricultural College. Ferdowsi University of Mashhad, Mashhad, I.R. Iran.
Soil and Crop Research and Development Center, Sainte-Foy, Quebec, Canada, G1V-2J3.
One hundred and four isolates of Bradyrhizobium japonicum were isolated from nodules of two different trap plants, Viz. Soya bean cultivars, Maple Glen and Orford which were inoculated with two different soilsamples (Ottawa and St-Hugus soils). All isolates were clustered based on PCR/RFLP of 16S-23S rRNAgenes. RFLP analysis was performed to characterize all the isolates using six different endonucleaseenzymes. The data was analyzed by using Jamp software. Using dendrogram data, all the isolates weregrouped into six different clusters. There were four and five clusters of Bradyrhizobium japonicum in Ottawaand St-Hugus soils, respectively. Three clusters were common between two cultivars of Soya bean wheninoculated with Ottawa soil and four common clusters were recognized when trap plants inoculated with St-Hugus soil. In Ottawa soil, cluster I was not detected by Orford cultivar, likewise in St-Hugus soil, cluster VIwas not detected by Maple Glen cultivar of Soya bean. Isolates of cluster III were dominantly trapped whenMaple Glen and Orford cultivars inoculated with Ottawa soil but isolates from clusters I, IV and III weretrapped when they were inoculated with St-Hugus soil. Since different cultivars trapped different isolate typesit can be concluded that for population studies of rhizobial bacteria different trap plants can provide a bettercomposition of native population of bacteria.
https://www.ijbiotech.com/article_6929_ce4b3934eaabe376de4a41946b68aa35.pdf
Bradyrhizobium japonicum
16S-23SrRNA
PCR/RFLP
Trap Plant
eng
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2004-04-01
2
2
97
100
6925
Interactive Effects of Heat Shock and Culture Density on Embryo Induction in Isolated Microspores Culture of Brassica napusL. cv. Global
Mohammad Reza Abdollahi
1
Ahmad Moieni
moieni_ahmad@yahoo.com
2
Mokhtar Jalali Javaran
m_jalali@modares.ac.ir
3
Plant Breeding Department, Faculty of Agriculture, Tarbiat Modarres University, P.O. Box: 14115-336, Tehran, I.R. Iran.
Plant Breeding Department, Faculty of Agriculture, Tarbiat Modarres University, P.O. Box: 14115-336, Tehran, I.R. Iran.
Plant Breeding Department, Faculty of Agriculture, Tarbiat Modarres University, P.O. Box: 14115-336, Tehran, I.R. Iran.
High yield and good quality embryos were obtained from cultures of isolated microspores of Brassica napus L. cv. Global. The donor plants were grown in a growth chamber at 15/10°C (day/night) with a 16/8hphotoperiod. Microspores were isolated from whole buds of 2.5-3.5 mm in length containing late-uninucleateand early-binucleate microspores. Different heat shock treatments including, 30°C for 10, 14 and 18days, 32°C for 2 and 3 days and 35°C for 18h followed by 30°C for 10 days and various culture densitiesincluding 60,000, 40,000 and 20,000 microspores per ml were used. Results showed significant differencesamong the heat shock treatments, the culture densities and their interaction for embryo induction. A large number of embryos were obtained from the microspores treated at 30°C for 18 days, 35°C for 18h followed by30°C for 10 days and 30°C for 14 days with a density of 60,000 microspores per ml.
https://www.ijbiotech.com/article_6925_90b9be945b3aedffa644f84741d6e50f.pdf
Brassica napus L
Microspore Culture
embryogenesis
Heat shock
Culture Density
eng
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2004-04-01
2
2
101
105
6903
Frequency of Bovine Lymphocyte Antigen DRB3.2 Alleles in Sarabi Cows
Fatemeh Montazer Torbati
1
Fereidoun Eftekhari Shahroudi
2
Mohammad Reza Nassiry
3
Abbas Safarnezhad
4
Mohadmmad Bagher Montazer Torbati
5
Department of Animal Science, Agriculture Faculty, Ferdowsi University, Mashhad, I.R. Iran.
Department of Animal Science, Agriculture Faculty, Ferdowsi University, Mashhad, I.R. Iran.
Department of Animal Science, Agriculture Faculty, Ferdowsi University, Mashhad, I.R. Iran.
Jahad-Keshavarzi Research Center, Mashhad, I.R. Iran.
Department of Animal Science, Agriculture Faculty, Tehran University, Tehran, I.R. Iran.
The second exon of the bovine Major Histocompatibility Complex (MHC) class II DRB3 gene was amplifiedby polymerase chain reaction (PCR) from 50 DNA samples of Iranian Sarabi cattle. Bovine DNA was isolatedfrom aliquots of whole blood. A two-step polymerase chain reaction followed by digestion withrestriction endonucleases RsaI, BstyI, and HaeIII was conducted on DNA from samples. Fifteen BovineLymphocyte Antigen (BoLA)-DRB3 alleles were assigned, including some that were only recentlydescribed for zebu cattle. Allelic frequencies ranged from 0.02 to 0.23. The most frequent alleles were *52(frequency = 0.23), *11 (0.18) and *23 (0.15). Results of this study demonstrate that the BoLA-DRB3.2 locusis highly polymorphic in Sarabi cattles.
https://www.ijbiotech.com/article_6903_48dfba01b11f8316ee80b19d8b420845.pdf
BoLA-DRB3.2
Polymerase chain reaction (PCR)
restriction fragment length polymorphism (RFLP)
Iranian Sarabi cows
eng
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2004-04-01
2
2
106
12
6926
A Simple Complexation Model and the Experimental Datafor Protein Extraction Using Reverse Micellar Systems
Ali Haghtalab
1
Shahriar Osfouri
2
Department of Chemical Engineering, Faculty of Engineering, Tarbiat Modarres University, P.O. Box 14115- 143, Tehran, IR Iran.
Department of Chemical Engineering, Faculty of Engineering, Tarbiat Modarres University, P.O. Box 14115- 143, Tehran, IR Iran.
and purification of proteins and enzymes in downstream processing. In this study a simple complexationmodel was developed for protein extraction using reverse micelles. We assumed that the size of proteinreverse micelle complex is a function of net charge of protein and salt concentration. The model has been applied to correlate the experimental data for reverse micellar extraction of bovine serum albumin (BSA) and lysozyme. The solutions of reverse micelles for extraction of BSA and lysozyme were composed ofcetyltrimethylammonium bromide (CTAB), a cationic surfactant, and sodium bis(2-ethylhexyl) phosphate(NaDEHP), an anionic surfactant, respectively. Moreover, the effects of surfactant concentration, pHof aqueous phase, and salt concentration were investigated. In comparison with experiment the results ofthe model for both systems are in very good agreement.
https://www.ijbiotech.com/article_6926_233d21c7233d912515fbf6ecdad0683e.pdf
Extraction
Modeling
Reverse Micelle
Bioseparation
BSA
Lysozyme
eng
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2004-04-01
2
2
113
122
6924
Fed-batch Cultivation of Recombinant Escherichia coli Producing Human Interferon-γ Under Controlled Specific Growth Rate
Rasoul Khalilzadeh
1
Seyed Abbas Shojaosadati
shoja@modares.ac.ir
2
Ali Bahrami
3
Nader Maghsoudi
4
Biotechnology Group, Chemical Engineering Department, Engineering Faculty, Tarbiat Modarres University, P.O. Box 14155-143, Tehran, I.R. Iran.
Biotechnology Group, Chemical Engineering Department, Engineering Faculty, Tarbiat Modarres University, P.O. Box 14155-143, Tehran, I.R. Iran.
Biotechnology Group, Chemical Engineering Department, Engineering Faculty, Tarbiat Modarres University, P.O. Box 14155-143, Tehran, I.R. Iran.
Neuroscience Research Center, Shaheed Beheshti University, P.O. Box: 19835- 181. Tehran, I.R. Iran.
A simple fed-batch process with pre-determined exponential feeding strategy for high-cell-density cultivationof recombinant E. coli BL21 (DE3) in defined medium was developed. In this feeding method glucose andglycerol were used as the sole sources of carbon and energy to increase the cell density exponentially atcontrolled specific growth rates, which do not cause the accumulation of acetate. Thus, sophisticated feedbackcontrol or extra equipment to prevent the accumulation of toxic level of acetate is not necessary. Thefinal cell densities of 100 and 118 gl-1 of dry cell mass for recombinant E. coli producing human interferon-γ(hIFN-γ) were obtained by using glucose and glycerol, respectively. The concentration of acetate was alwaysmaintained below toxic level. The specific yield of hIFN-γ with glucose and glycerol was 93 and 92 mgg-1of dry cell mass, and the overall productivity of hIFN-γ was 0.16 and 0.14 gl-1h-1 for these two carbon sources, respectively.
https://www.ijbiotech.com/article_6924_9108db3709b7ac963427a974bbfd0c09.pdf
Human interferon-gamma
Fed-batch cultivation
recombinant E. coli
Specific growth rate
Glucose
Glycerol
eng
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2004-04-01
2
2
123
131
6923
Cloning and Expression of the Heterogenic Vacuolating Cytotoxin From an Iranian Helicobacter pylori Strain
Yeganeh Talebkhan
1
Fereidoun Mahboudi
mahboudi@pasteur.ac.ir
2
Ramin Sarrami
3
Farzaneh Barkhordari
fbarkhordari@yahoo.com
4
Mehrnaz Amani
5
Marjan Mohammadi
6
Biotechnology Research Center, Pasteur Institute of Iran and National Biotechnology Network, Tehran, I.R.Iran.
Biotechnology Research Center, Pasteur Institute of Iran and National Biotechnology Network, Tehran, I.R.Iran.
Biotechnology Research Center, Pasteur Institute of Iran and National Biotechnology Network, Tehran, I.R.Iran.
Biotechnology Research Center, Pasteur Institute of Iran and National Biotechnology Network, Tehran, I.R.Iran.
Biotechnology Research Center, Pasteur Institute of Iran and National Biotechnology Network, Tehran, I.R.Iran.
Biotechnology Research Center, Pasteur Institute of Iran and National Biotechnology Network, Tehran, I.R.Iran.
Several reports indicate that the nonconserved genes of Helicobacter pylori (H. pylori) in particular its cytotoxinare widely heterogeneous among various geographic locations and this is manifested at the proteinlevel ranging from 5-15% which demands access to locally deduced protein antigens for inclusion intodiagnostic kits and/or inclusion as a vaccine component for the target population. We have previouslydemonstrated such variations via PCR-RFLP analysis between Iranian and western H. pylori strains. vacAgene from a selected strain of the most prevalent RFLP category among Iranian strains, was partiallysequenced which revealed 8.3% dissimilarity with reference strains at protein level. This drastic differenceprompted us to subclone the vacA coding region into an expression vector to produce the recombinant protein.Full sequencing of the coding region demonstrated 8-9% amino acid difference with American andGerman reference strains. Recombinant protein expression yielded 4% of the total E. coli proteins.Histidine tag allowed for purification of the recombinant VacA using immobilized metal affinity chromatography(IMAC). Identity of the recombinant protein was repeatedly confirmed by Western blot analysis usingpatient serum, rabbit hyper immune serum as well as anti-His monoclonal antibody.
https://www.ijbiotech.com/article_6923_845c57ad87d557a4b626f48da447c907.pdf
Helicobacter pylori
Escherichia coli
Recombinant
cytotoxin
vacA
heterogeneity
Cloning
Expression
Purification
eng
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2004-04-01
2
2
132
135
6904
Carrier Determination in a Hemophilia B Family Using Single Strand Conformation Polymorphism (SSCP) and Sequencing
Morteza Karimipoor
1
Sirous Zeinali
sirous.zeinali@ut.ac.ir
2
Reza Safaee
3
Manijheh Lak
4
Nafiseh Nafissi
5
Biotechnology Research Center, Pasteur Institute of Iran, Tehran, 2Hemophilia Center, Imam Khomeini Hospital, Tehran, I.R. Iran.
Biotechnology Research Center, Pasteur Institute of Iran, Tehran, 2Hemophilia Center, Imam Khomeini Hospital, Tehran, I.R. Iran.
Hemophilia Center, Imam Khomeini Hospital, Tehran, I.R. Iran.
Hemophilia Center, Imam Khomeini Hospital, Tehran, I.R. Iran.
Biotechnology Research Center, Pasteur Institute of Iran, Tehran, I.R. IRan.
Hemophilia B is an X-linked recessive bleeding disorder caused by heterogeneous mutations in factor IXgene. In about one-third of cases it arises by a new mutation in germ-line cells. In this study carrier testingwas performed for females of a family with only one affected individual by single strand conformation polymorphism (SSCP). Results indicated that the SSCP band shift in the propositus was de novo and his mother and also sisters were not carrier. This finding was also confirmed by sequencing.
https://www.ijbiotech.com/article_6904_399148894279b5e60e29f9e4f6a932aa.pdf
Hemophilia B
Mutation
SSCP
Carrier Testing