National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2
2
2004
04
01
Endophytic Fungi, Characteristics and Their Potential for Genetic Manipulation
75
83
EN
Aghafakhr
Mirlohi
Department of Plant Breeding, Isfahan University of Technology, Isfahan, Iran.
Mohammad Reza
Sabzalian
Department of Plant Breeding, Isfahan University of Technology, Isfahan, Iran.
Mojtaba
Khayyam Nekouei
Agriculture Biotechnology Institute
of Central Region of I.R. Iran.
Unique benign endophytes from Ascomycets have wide distribution among grass species. The symbiotic<br />fungi enhance plant characters including performance, insect and mammalian deterrence, nematode resistance and tolerance to drought, salt and other biotic and abiotic stresses. Endophytes from genus<br />Neotyphodium (Acremonium) are of the major focus than their ancestors, and Epichloe species, because<br />the formers have lost their sexual reproduction. Therefore they should be genetically stable, and most<br />importantly, they cannot disassociate from host tissues, and are transferred vertically. They are maternally<br />inherited and are therefore attractive for genetic transformation without the concern about gene<br />escape. Some marker genes have been successfully transferred to endophyte Neotyphodium coenophialum<br />and Neotyphodium lolii existing in Festuca arundinacea Schreb. and Lolium perenne L., respectively.<br />Furthermore, gene silencing has been proved to be feasible for eliminating traits, which are economically<br />harmful. Methods of direct DNA uptake using polyethylene glycol (PEG) and electroporation have been<br />found to be useful in transformation of these fungi. Transgenic fungi can be reinserted into the host without<br />need 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.
Endophyte,Molecular marker,Neotyphodium,Transformation
https://www.ijbiotech.com/article_6935.html
https://www.ijbiotech.com/article_6935_724bf7d6beddf379db85387c7ad1e249.pdf
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2
2
2004
04
01
Cloning and Expression of the Coat Protein Gene of Barley Yellow Dwarf Virus-PAV in Escherichia coli
84
89
EN
Masoud
Shams-bakhsh
0000-0003-2923-2668
Department of Plant Pathology, Tarbiat Modarres University, P.O. Box: 14115-143, Tehran, I.R. Iran.
shamsbakhsh@modares.ac.ir
Robert Henry
Symons
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 viral<br />coat protein without purifying the virus. To produce recombinant coat protein, the coding sequence was<br />first amplified from a PAV full-length cDNA clone by polymerase chain reaction (PCR), ligated into a vector<br />(pBluescript SK+) to check the sequence, and subcloned into an expression vector (pGEX-2T). It was<br />then transformed into Escherichia coli DH5α by electroporation. The open reading frame 3 (ORF3) was<br />linked in-frame to the gene encoding glutathione-Stransferase (GST; 26 kDa) and expression induced by<br />IPTG. The expressed coat protein was purified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis<br />(SDS-PAGE) for use as an immunogen. The antisera to BYDV-PAV recombinant coat protein reacted in<br />Western blot analysis with partially purified BYDV-PAV. These antisera were also used to detect BYDV-PAV by<br />immunogold electron microscopy of thin section of barley tissues. The results indicated that BYDV-PAV coat<br />protein can be produced in high yields by E. coli, which provides the ability of simple purification, and because<br />of proper antigencity, can be exploited for diagnostic applications.
Barley yellow dwarf virus,Expression vector,polyclonal antibodies,E. coli,Coat protein,PAV
https://www.ijbiotech.com/article_6928.html
https://www.ijbiotech.com/article_6928_29ead82d79f50238f1f3877b5df856e4.pdf
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2
2
2004
04
01
The Effect of Trap Plants on the Population Diversity of Bradyrhizobium japonicum
90
96
EN
Amir
Lakzian
Soil Science Department. Agricultural College. Ferdowsi University of Mashhad, Mashhad, I.R. Iran.
alakzian@yahoo.com
Eden
Bromfield
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 soil<br />samples (Ottawa and St-Hugus soils). All isolates were clustered based on PCR/RFLP of 16S-23S rRNA<br />genes. RFLP analysis was performed to characterize all the isolates using six different endonuclease<br />enzymes. The data was analyzed by using Jamp software. Using dendrogram data, all the isolates were<br />grouped into six different clusters. There were four and five clusters of Bradyrhizobium japonicum in Ottawa<br />and St-Hugus soils, respectively. Three clusters were common between two cultivars of Soya bean when<br />inoculated with Ottawa soil and four common clusters were recognized when trap plants inoculated with St-<br />Hugus soil. In Ottawa soil, cluster I was not detected by Orford cultivar, likewise in St-Hugus soil, cluster VI<br />was not detected by Maple Glen cultivar of Soya bean. Isolates of cluster III were dominantly trapped when<br />Maple Glen and Orford cultivars inoculated with Ottawa soil but isolates from clusters I, IV and III were<br />trapped when they were inoculated with St-Hugus soil. Since different cultivars trapped different isolate types<br />it can be concluded that for population studies of rhizobial bacteria different trap plants can provide a better<br />composition of native population of bacteria.
Bradyrhizobium japonicum,16S-23SrRNA,PCR/RFLP,Trap Plant
https://www.ijbiotech.com/article_6929.html
https://www.ijbiotech.com/article_6929_ce4b3934eaabe376de4a41946b68aa35.pdf
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2
2
2004
04
01
Interactive Effects of Heat Shock and Culture Density on Embryo Induction in Isolated Microspores Culture of Brassica napusL. cv. Global
97
100
EN
Mohammad Reza
Abdollahi
Plant Breeding Department, Faculty of Agriculture, Tarbiat Modarres University, P.O. Box: 14115-336, Tehran,
I.R. Iran.
Ahmad
Moieni
Plant Breeding Department, Faculty of Agriculture, Tarbiat Modarres University, P.O. Box: 14115-336, Tehran,
I.R. Iran.
moieni_ahmad@yahoo.com
Mokhtar
Jalali Javaran
0000-0002-4668-0764
Plant Breeding Department, Faculty of Agriculture, Tarbiat Modarres University, P.O. Box: 14115-336, Tehran,
I.R. Iran.
m_jalali@modares.ac.ir
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 16/8h<br />photoperiod. Microspores were isolated from whole buds of 2.5-3.5 mm in length containing late-uninucleate<br />and early-binucleate microspores. Different heat shock treatments including, 30°C for 10, 14 and 18<br />days, 32°C for 2 and 3 days and 35°C for 18h followed by 30°C for 10 days and various culture densities<br />including 60,000, 40,000 and 20,000 microspores per ml were used. Results showed significant differences<br />among 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 by<br />30°C for 10 days and 30°C for 14 days with a density of 60,000 microspores per ml.<br /><br />
Brassica napus L,Microspore Culture,embryogenesis,Heat shock,Culture Density
https://www.ijbiotech.com/article_6925.html
https://www.ijbiotech.com/article_6925_90b9be945b3aedffa644f84741d6e50f.pdf
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2
2
2004
04
01
Frequency of Bovine Lymphocyte Antigen DRB3.2 Alleles in Sarabi Cows
101
105
EN
Fatemeh
Montazer Torbati
Department of Animal Science, Agriculture Faculty, Ferdowsi University, Mashhad, I.R. Iran.
Fereidoun
Eftekhari Shahroudi
Department of Animal Science, Agriculture Faculty, Ferdowsi University, Mashhad, I.R. Iran.
Mohammad Reza
Nassiry
Department of Animal Science, Agriculture Faculty, Ferdowsi University, Mashhad, I.R. Iran.
Abbas
Safarnezhad
Jahad-Keshavarzi Research Center, Mashhad, I.R. Iran.
Mohadmmad Bagher
Montazer Torbati
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 amplified<br />by polymerase chain reaction (PCR) from 50 DNA samples of Iranian Sarabi cattle. Bovine DNA was isolated<br />from aliquots of whole blood. A two-step polymerase chain reaction followed by digestion with<br />restriction endonucleases RsaI, BstyI, and HaeIII was conducted on DNA from samples. Fifteen Bovine<br />Lymphocyte Antigen (BoLA)-DRB3 alleles were assigned, including some that were only recently<br />described for zebu cattle. Allelic frequencies ranged from 0.02 to 0.23. The most frequent alleles were *52<br />(frequency = 0.23), *11 (0.18) and *23 (0.15). Results of this study demonstrate that the BoLA-DRB3.2 locus<br />is highly polymorphic in Sarabi cattles.<br /><br />
BoLA-DRB3.2,Polymerase chain reaction (PCR),restriction fragment length polymorphism (RFLP),Iranian Sarabi cows
https://www.ijbiotech.com/article_6903.html
https://www.ijbiotech.com/article_6903_48dfba01b11f8316ee80b19d8b420845.pdf
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2
2
2004
04
01
A Simple Complexation Model and the Experimental Datafor Protein Extraction Using Reverse Micellar Systems
106
12
EN
Ali
Haghtalab
Department of Chemical Engineering, Faculty of Engineering, Tarbiat Modarres University, P.O. Box 14115- 143, Tehran, IR Iran.
Shahriar
Osfouri
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 complexation<br />model 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 of<br />cetyltrimethylammonium bromide (CTAB), a cationic surfactant, and sodium bis(2-ethylhexyl) phosphate<br />(NaDEHP), an anionic surfactant, respectively. Moreover, the effects of surfactant concentration, pH<br />of aqueous phase, and salt concentration were investigated. In comparison with experiment the results of<br />the model for both systems are in very good agreement.<br /><br />
Extraction,Modeling,Reverse Micelle,Bioseparation,BSA,Lysozyme
https://www.ijbiotech.com/article_6926.html
https://www.ijbiotech.com/article_6926_233d21c7233d912515fbf6ecdad0683e.pdf
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2
2
2004
04
01
Fed-batch Cultivation of Recombinant Escherichia coli Producing Human Interferon-γ Under Controlled Specific Growth Rate
113
122
EN
Rasoul
Khalilzadeh
Biotechnology Group, Chemical Engineering Department, Engineering Faculty, Tarbiat Modarres University,
P.O. Box 14155-143, Tehran, I.R. Iran.
Seyed Abbas
Shojaosadati
Biotechnology Group, Chemical Engineering Department, Engineering Faculty, Tarbiat Modarres University,
P.O. Box 14155-143, Tehran, I.R. Iran.
shoja@modares.ac.ir
Ali
Bahrami
Biotechnology Group, Chemical Engineering Department, Engineering Faculty, Tarbiat Modarres University,
P.O. Box 14155-143, Tehran, I.R. Iran.
Nader
Maghsoudi
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 cultivation<br />of recombinant E. coli BL21 (DE3) in defined medium was developed. In this feeding method glucose and<br />glycerol were used as the sole sources of carbon and energy to increase the cell density exponentially at<br />controlled specific growth rates, which do not cause the accumulation of acetate. Thus, sophisticated feedback<br />control or extra equipment to prevent the accumulation of toxic level of acetate is not necessary. The<br />final cell densities of 100 and 118 gl-1 of dry cell mass for recombinant E. coli producing human interferon-γ<br />(hIFN-γ) were obtained by using glucose and glycerol, respectively. The concentration of acetate was always<br />maintained below toxic level. The specific yield of hIFN-γ with glucose and glycerol was 93 and 92 mgg-1<br />of 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.<br /><br />
Human interferon-gamma,Fed-batch cultivation,recombinant E. coli,Specific growth rate,Glucose,Glycerol
https://www.ijbiotech.com/article_6924.html
https://www.ijbiotech.com/article_6924_9108db3709b7ac963427a974bbfd0c09.pdf
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2
2
2004
04
01
Cloning and Expression of the Heterogenic Vacuolating Cytotoxin From an Iranian Helicobacter pylori Strain
123
131
EN
Yeganeh
Talebkhan
Biotechnology Research Center, Pasteur Institute of Iran and National Biotechnology Network, Tehran, I.R.Iran.
Fereidoun
Mahboudi
0000-0002-3380-5257
Biotechnology Research Center, Pasteur Institute of Iran and National Biotechnology Network, Tehran, I.R.Iran.
mahboudi@pasteur.ac.ir
Ramin
Sarrami
Biotechnology Research Center, Pasteur Institute of Iran and National Biotechnology Network, Tehran, I.R.Iran.
Farzaneh
Barkhordari
Biotechnology Research Center, Pasteur Institute of Iran and National Biotechnology Network, Tehran, I.R.Iran.
fbarkhordari@yahoo.com
Mehrnaz
Amani
Biotechnology Research Center, Pasteur Institute of Iran and National Biotechnology Network, Tehran, I.R.Iran.
Marjan
Mohammadi
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 cytotoxin<br />are widely heterogeneous among various geographic locations and this is manifested at the protein<br />level ranging from 5-15% which demands access to locally deduced protein antigens for inclusion into<br />diagnostic kits and/or inclusion as a vaccine component for the target population. We have previously<br />demonstrated such variations via PCR-RFLP analysis between Iranian and western H. pylori strains. vacA<br />gene from a selected strain of the most prevalent RFLP category among Iranian strains, was partially<br />sequenced which revealed 8.3% dissimilarity with reference strains at protein level. This drastic difference<br />prompted us to subclone the vacA coding region into an expression vector to produce the recombinant protein.<br />Full sequencing of the coding region demonstrated 8-9% amino acid difference with American and<br />German reference strains. Recombinant protein expression yielded 4% of the total E. coli proteins.<br />Histidine tag allowed for purification of the recombinant VacA using immobilized metal affinity chromatography<br />(IMAC). Identity of the recombinant protein was repeatedly confirmed by Western blot analysis using<br />patient serum, rabbit hyper immune serum as well as anti-His monoclonal antibody.<br /><br />
Helicobacter pylori,Escherichia coli,Recombinant,cytotoxin,vacA,heterogeneity,Cloning,Expression,Purification
https://www.ijbiotech.com/article_6923.html
https://www.ijbiotech.com/article_6923_845c57ad87d557a4b626f48da447c907.pdf
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2
2
2004
04
01
Carrier Determination in a Hemophilia B Family Using Single Strand Conformation Polymorphism (SSCP) and Sequencing
132
135
EN
Morteza
Karimipoor
Biotechnology Research Center, Pasteur Institute of Iran, Tehran, 2Hemophilia Center, Imam Khomeini Hospital, Tehran, I.R. Iran.
Sirous
Zeinali
Biotechnology Research Center, Pasteur Institute of Iran, Tehran, 2Hemophilia Center, Imam Khomeini Hospital, Tehran, I.R. Iran.
sirous.zeinali@ut.ac.ir
Reza
Safaee
Hemophilia Center, Imam Khomeini
Hospital, Tehran, I.R. Iran.
Manijheh
Lak
Hemophilia Center, Imam Khomeini
Hospital, Tehran, I.R. Iran.
Nafiseh
Nafissi
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 IX<br />gene. In about one-third of cases it arises by a new mutation in germ-line cells. In this study carrier testing<br />was 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.<br /><br />
Hemophilia B,Mutation,SSCP,Carrier Testing
https://www.ijbiotech.com/article_6904.html
https://www.ijbiotech.com/article_6904_399148894279b5e60e29f9e4f6a932aa.pdf