National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2
3
2004
07
01
The Primed In Situ (PRINS) Technique: An Alternative Approach for Preimplantation Chromosomal Diagnosis
149
157
EN
Hossein
Mozdarani
Deptartment of Medical Genetics, School of Medical Sciences, Tarbiat Modarres Univ., Tehran, I.R. Iran.
Franck
Pellestor
2Institute of Human Genetics, CNRS UPR ll42, Montpellier, France.
Preimplantation genetic diagnosis (PGD) is a novel approach for the prevention of genetic disorders in couples at risk of having offspring with genetic disease. Although the original idea dates back to early 1960s when sexing of rabbit blastocysts was attempted, the first clinical application of PGD was reported about three decades later, describing the use of PCR for sexing embryos from couples at risk of X-linked disease. The development of PCR-based tests led to PGD for screening well known monogenic diseases such as thalassaemia and cystic fibrosis. The introduction of fluorescence in situ hybridization (FISH) quickly replaced PCR-based methods which had led to misdiagnoses for sexing embryos. FISH can be used for aneuploidy screening of up to seven clinically significant chromosomes and translocation detection. The advent of molecular genetic techniques has brought forth new procedures for in situ chromosomal analysis. One of these techniques is the primed in situ labeling (PRINS) procedure which constitutes a fast and efficient alternative to conventional fluorescence in situ hybridization for nucleic acid detection. This technique has the potential to become a powerful tool for cytogenetic investigations. The recent achievements reported show that PRINS can constitute an efficient complement to PCR and FISH. Adaptation of this technique to preimplantation embryo screening both at chromosomal level and gene localization opens a promising perspective for being used in the field of PGD.<br /><br />
Preimplantation genetic diagnosis,PCR,fish,PRINS,chromosomal abnormality
https://www.ijbiotech.com/article_6915.html
https://www.ijbiotech.com/article_6915_59f6c82e1269ed50bf8c13469895396a.pdf
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2
3
2004
07
01
Development of Transgenic Mice Harboring Ovine Beta Lactoglobulin-Calcitonin Transgene
158
163
EN
Ahmadreza
Niavarani
Department of Biotechnology, Pasteur Institute of Iran, Tehran, I.R. Iran.
Sirous
Zeinali
Department of Biotechnology, Pasteur Institute of Iran, Tehran, I.R. Iran.
sirous.zeinali@ut.ac.ir
Mohsen
Karimi
Department of Biotechnology, Pasteur Institute of Iran, Tehran, I.R. Iran.
Minoo
Rassoulzadegan
INSERM U636, University of
Nice-Sophia Antipolis, France.
Expression of foreign proteins in mammalian milk is becoming a widespread strategy for high-level production<br />of recombinant pharmaceuticals, especially those with the most complex post-translational modifications.<br />A gene construct was generated, consisting of 10.7 kbp of the ovine beta-lactoglobulin (oBLG) gene<br />including its promoter and 3´ flanking region with the calcitonin coding sequences inserted in-frame into the<br />oBLG fifth exon. The gene construct was purified using CsCl gradient, released from vector, and gel-purified. It<br />was microinjected into fertilized mouse oocytes. These oocytes were then transferred to pseudo-pregnant foster mice. The pups born from foster mice were genotyped using PCR, slot blot, and Southern blot techniques. Among 9 mice which showed positive PCR results, only 6 mice transmitted the transgene to the<br />next generation. Therefore, 6 transgenic lines were established which stably transmitted their transgene to<br />their progeny.
transgenic mice,Milk,Calcitonin,Beta-lactoglobulin
https://www.ijbiotech.com/article_6936.html
https://www.ijbiotech.com/article_6936_480e6518a88c715b47a4425199803d3a.pdf
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2
3
2004
07
01
Development of PCR-ELISA Technique for Determination of HLADRB1*01 Group Alleles
164
169
EN
Habib
Nasiri
Department of Medical Biotechnology, Faculty of Medical Science, Tarbiat Modarres University, Tehran, I.R. Iran.
Mehdi
Forouzandeh
Department of Medical Biotechnology, Faculty of Medical Science, Tarbiat Modarres University, Tehran, I.R. Iran.
mehdi.forouzandeh@gmail.com
Aliakbar
Pourfathollah
Department of Hematology, Faculty of Medical Science, Tarbiat Modarres University, Tehran, I.R. Iran.
Mohammad Javad
Rasaee
Department of Medical Biotechnology, Faculty of Medical Science, Tarbiat Modarres University, Tehran, I.R. Iran.
Fatemeh
Rahbarizadeh
Department of Medical Biochemistry,
Faculty of Medical Science, Tarbiat Modarres University, Tehran, I.R. Iran.
rahbarif@modares.ac.ir
We have developed an allotyping assay for detection of four HLA DRB1*01 group alleles based on polymerase<br />chain reaction and solution hybridization in a microtiter plate. Using group specific primers a region<br />within exon 2 of HLA DRB1 gene was amplified by PCR. Labeling of PCR product was achieved by<br />adding small amount of Dig-dUTP in place of dTTP. Labeled PCR product was hybridized to allele (HLA<br />DRB1*0101, *0102, *0103 and *0104) specific and a group (HLA DRB1*01) specific oligonucleotide probes<br />in separate wells of the plate. Hybridized amplicones were detected by an enzymatic procedure. Ninety<br />DNA samples were tested in parallel with PCR-SSP typing. The results were found to be well correlated by<br />two methods. These results further suggest that, PCRELISA would be a rapid, specific and simple method<br />that can be used for high resolution HLA typing before bone marrow and stem cell transplantation.<br /><br />
HLA DRB1,PCR-ELISA,Hybridization,Group specific amplification
https://www.ijbiotech.com/article_6905.html
https://www.ijbiotech.com/article_6905_51cd755ebf5d6cc0573fefc000d8a51f.pdf
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2
3
2004
07
01
High-level expression and evaluation of the antigenicity of a recombinant Toxoplasma gondii GRA2 Protein
170
176
EN
Majid
Golkar
Department of Parasitology, Pasteur Institute of Iran, Tehran, I.R. Iran.
Sima
Rafati
Department of Immunology, Pasteur Institute of Iran, Tehran, I.R. Iran.
Yasaman
Taslimi
Department of Immunology, Pasteur Institute of Iran, Tehran, I.R. Iran.
Tahereh
Taheri
Department of Immunology, Pasteur Institute of Iran, Tehran, I.R. Iran.
Fatemeh
Doustdari
Department of Immunology, Pasteur Institute of Iran, Tehran, I.R. Iran.
Mehdi
Assmar
Department of Parasitology, Pasteur Institute of Iran, Tehran, I.R. Iran.
asmar@institute.pasteur.ac.ir
Toxoplasmosis is a worldwide infection which is commonly asymptomatic but can cause serious health problems in immunocompromised individuals and fetus. GRA2 is a dense granule protein of Toxoplasma<br />gondii, which induces strong antibody and T-cell response in human and mice. The purpose of this<br />study was to prepare recombinant GRA2 and evaluate its antigenic properties using infected mice sera. To<br />reach this point, GRA2 gene was highly expressed as a fusion protein with Thioredoxin (TRX) in Escherichia<br />coli BL21pLysS strain. The protein was purified in a single step on Ni-NTA affinity column. TRX-GRA2 was<br />confirmed by Western blot technique using a monoclonal antibody specific for GRA2. Sera from mice<br />infected with Toxoplasma gondii showed high reactivity toward GRA2 and the level of IgG2a isotype was<br />predominant and significantly higher than IgG1.Taken together, TRX-GRA2 might be considered as an ideal<br />antigen to be used as a vaccine target as well as diagnostic tool for detection of toxoplasmosis.<br /><br />
Toxoplasma gondii,GRA2,Thioredoxin,E. coli,Expression
https://www.ijbiotech.com/article_6907.html
https://www.ijbiotech.com/article_6907_157af281291b57b891b85b3d95a6f140.pdf
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2
3
2004
07
01
Disruption of GAL1 Gene in Saccharomyces cerevisiae Leads to Higher Ethanol Production
177
82
EN
Abbas
Rezaee
Faculty of Medical Sciences, Tarbiat Modarres University, Tehran, I.R. Iran,
Hyun Ah
Kang
Microbial Metabolic Engineering
Research Units, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Taejon, South Korea.
Sang Ki
Rhee
Microbial Metabolic Engineering
Research Units, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Taejon, South Korea.
GAL gene family is a set of structural and regulatory genes that enables cells to utilize galactose as a carbon<br />source in Saccharomyces cerevisiae. Phosphorylation of intracellular galactose can be catalyzed by<br />galactokinase (encoded by GAL1 gene). In this study role of GAL1 gene on ethanol production by S. cerevisiae<br />together with physiological characterization of GAL1 mutant strain was studied. Aerobic cultivation was carried out with wild-type strain and the GAL1 mutant. The GAL1 mutant strain displayed fermentative growth in early exponential phase. Deletion of the GAL1 gene was shown to have a major impact on biomass and ethanol formation. The GAL1 mutant exhibited a decrease in growth rate and increased ethanol production. Furthermore the results showed that glucose consumption by GAL1 mutants did not favor biomass formation, rather cause excessive respiro-fermentative metabolism, with whereas could linear increase in ethanol production.<br /><br />
GAL1,Saccharomyces Cerevisiae,Gene disruption,Ethanol
https://www.ijbiotech.com/article_6908.html
https://www.ijbiotech.com/article_6908_8ffa66790224dab6eb3d7b2061bef2b4.pdf
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2
3
2004
07
01
Cloning, expression and purification of Clostridium botulinum neurotoxin type E binding domain
183
188
EN
Mir Latif
Mousavi
Department of Biology, Faculty of Science, Imam Hossain University, Tehran, I.R. Iran.
Shideh
Montaser Kouhsari
Department of Biology, Faculty of Science, Tehran University, Tehran, I.R. Iran.
Shahram
Nazarian
Department of Biology, Faculty of Science, Imam Hossain University, Tehran, I.R. Iran.
shahramnazarian@yahoo.com
Iraj
Rasooli
Department of Biology, College of Basic Science, Shahed University, Tehran, I.R. Iran.
irasooli@yahoo.com
Jafar
Amani
Department of Biology, Faculty of Science, Imam Hossain University, Tehran, I.R. Iran.
afar.amani@gmail.com
Botulinum neurotoxins constitute a family of bacterial toxins for botulism syndrome in human. The toxins<br />bind with high affinity to nerve cells where they cause a complete inhibition and release of neurotransmitters<br />and thereby produce flaccid paralysis. In this study the binding domain of type E neurotoxin was isolated by<br />PCR and expressed in a proper expression vector. The results of this investigation can be used as a tool<br />to study the mechanism of binding of holotoxins. This study is also implicated in antibody production against<br />botulism syndrome.
Botulinum neurotoxin type E,Binding Domain,Expression
https://www.ijbiotech.com/article_6909.html
https://www.ijbiotech.com/article_6909_79f3057bec43722c922ee415a34c6977.pdf
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2
3
2004
07
01
Purification of tyrosinase from edible mushroom
189
194
EN
Kamahldin
Haghbeen
0000-0003-3011-5629
Department of Biochemistry, National Research Institute for Genetic Engineering and Biotechnology, Tehran, I.R. Iran.
kamahl@nigeb.ac.ir
Ferdous
Rastgar Jazii
Department of Biochemistry, National Research Institute for Genetic Engineering and Biotechnology, Tehran, I.R. Iran.
rastgarjazii@gmail.com
Ali Asghar
Karkhane
Department of Biochemistry, National Research Institute for Genetic Engineering and Biotechnology, Tehran, I.R. Iran.
karkhane@nigeb.ac.ir
Shahrzad
Shareefi Borojerdi
Department of Biochemistry, National Research Institute for Genetic Engineering and Biotechnology, Tehran, I.R. Iran.
A simple preparative method was developed for purification of Tyrosinase from edible mushroom (Agaricus<br />bispora). A homogenized extract of mushroom was first saturated by ammonium sulfate. The desired precipitate was mixed thoroughly with DEAE-Cellulose (DE-52) and washed out to produce melanin free precipitate. The obtained protein solution was dialyzed against running water for 4 hrs, then, concentrated and<br />chromatographed on a DE-52 column. On the basis of the activities assay, the eluted fractions by 150 mM<br />salt solution were selected for further purification. The collected fractions were pooled and chromatographed<br />on a Sephadex G-200 column. Polyacrylamide gel electrophoresis (PAGE) of the purified tyrosinase produced<br />a single band right beside the commercial sample obtained from Sigma Company at 128 kDa. The<br />lyophilized form of the purified Tyrosinase had a purification degree of 104 and showed strong cresolase<br />and catecholase activities when compared to a commerically available tyrosinase.<br /><br />
Tyrosinase,Edible mushroom,Purification,Extraction
https://www.ijbiotech.com/article_6910.html
https://www.ijbiotech.com/article_6910_2ee60dd3ea3eee72653e5c43423e3bb7.pdf
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2
3
2004
07
01
Comparative analyses of the genetic diversity among bread wheat genotypes based on RAPD and SSR markers
195
202
EN
Mohammad Reza
Naghavi
0000-0002-2427-3332
Plant Breeding Department, Faculty of Agriculture, University of Tehran, Karaj, I.R. Iran.
mnaghavi@ut.ac.ir
Mohsen
Mardi
The Agricultural Biotechnology Research Institute of Iran. Karaj, I.R. Iran.
mardi@abrii.ac.ir
Hossein Ali
Ramshini
Plant Breeding Department, Faculty of Agriculture, University of Tehran, Karaj, I.R. Iran.
Bahman
Fazelinasab
Plant Breeding Department, Faculty of Agriculture, University of Tehran, Karaj, I.R. Iran.
Two different DNA-based techniques viz, randomly amplified polymorphic DNA (RAPD) and simple sequence repeat (SSR) markers were used to estimate genetic diversity among bread wheat. A total of 188 clear and repeatable bands were amplified from 17 selected RAPD primers, and 101 fragments were detected from 35 SSR primer pairs. The level of polymorphism was 88% with RAPDs compared to 100% with SSRs. Mean genetic similarity was estimated to be 0.88 based on RAPDs and 0.85 using SSRs. The wide range of genetic similarity was obtained by SSR than RAPD, reflecting the hypervariability of SSR markers and their high resolution power. Matrix correlation analyses suggested that a good representation of the relationships among the bread wheat<br />cultivars/lines can be obtained by using RAPDs alone or in combination with SSRs, but SSRs alone cannot<br />be used for this purpose. Both techniques discriminated the genotypes very effectively. On the hand,<br />RAPDs were able to discriminate the cultivars Alvand and Ghods, whereas the cultivars Sardari and Ghods<br />were discriminated only by SSRs. The use of PCRbased assays having advantage of being quick, easy<br />to use and refractory to many environmental influences can complement traditional methods of germplasm characterization.
Wheat,Diversity,RAPD,SSR
https://www.ijbiotech.com/article_6911.html
https://www.ijbiotech.com/article_6911_dcbb61f4f6658e6cf7e07f63f45aa71b.pdf
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2
3
2004
07
01
Detection of NAD(P)H: quinone oxidoreductase 609C T polymorphism in blood and archival human tissues using a simple PCR method
203
206
EN
Firouzeh
Biramijamal
National Institute for Genetic Engineering and Biotechnology (NIGEB), Tehran, I.R.Iran.
Mohammed Hossein
Sanati
National Institute for Genetic Engineering and Biotechnology (NIGEB), Tehran, I.R.Iran.
Guity
Iravanloo
Department of Pathology, Cancer Institute of Tehran, Tehran University of Medical Sciences, Tehran, I.R.Iran.
Kourosh
Shamimi
Department of Surgery, Tehran
University of Medical Sciences, Tehran, I.R.Iran.
Dariush D.
Farhud
Genetic Clinic Vallieasr square, Tehran, I.R. Iran.
NAD(P)H: quinone oxidoreductase (NQO1) plays an important role in detoxification of numerous endogenous<br />and foreign compounds. This gene has a single nucleotide polymorphism at site of codon 187<br />(CCT TCT). Recently, it has been demonstrated that individuals with T allele may exhibit resistance to<br />quinone based anticancer drugs such as mitomycin C. In the present study, a simple and feasible method was<br />developed for detection of NQO1 genotype. In this modified procedure, dimethylsulfoxide (DMSO) and<br />Triton X-100 were eliminated, also, PCR cycling conditions were modified to improve the PCR products from<br />blood and formalin-fixed, paraffin-embedded tissues. PCR-RFLP and DNA sequencing analysis carried out<br />on a limited number of blood and archival samples. It is suggested that this procedure convenient for NQO1<br />genotyping.
NQO1,Polymorphism,simple PCR-RFLP method
https://www.ijbiotech.com/article_6912.html
https://www.ijbiotech.com/article_6912_8474984e4ad22a9ff1c3413f536381e3.pdf