2024-03-28T23:19:07Z
https://www.ijbiotech.com/?_action=export&rf=summon&issue=1155
Iranian Journal of Biotechnology
IRAN J BIOTECH
1728-3043
1728-3043
2004
2
3
The Primed In Situ (PRINS) Technique: An Alternative Approach for Preimplantation Chromosomal Diagnosis
Hossein
Mozdarani
Franck
Pellestor
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.
Preimplantation genetic diagnosis
PCR
fish
PRINS
chromosomal abnormality
2004
07
01
149
157
https://www.ijbiotech.com/article_6915_59f6c82e1269ed50bf8c13469895396a.pdf
Iranian Journal of Biotechnology
IRAN J BIOTECH
1728-3043
1728-3043
2004
2
3
Development of Transgenic Mice Harboring Ovine Beta Lactoglobulin-Calcitonin Transgene
Ahmadreza
Niavarani
Sirous
Zeinali
Mohsen
Karimi
Minoo
Rassoulzadegan
Expression of foreign proteins in mammalian milk is becoming a widespread strategy for high-level productionof recombinant pharmaceuticals, especially those with the most complex post-translational modifications.A gene construct was generated, consisting of 10.7 kbp of the ovine beta-lactoglobulin (oBLG) geneincluding its promoter and 3´ flanking region with the calcitonin coding sequences inserted in-frame into theoBLG fifth exon. The gene construct was purified using CsCl gradient, released from vector, and gel-purified. Itwas 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 thenext generation. Therefore, 6 transgenic lines were established which stably transmitted their transgene totheir progeny.
transgenic mice
Milk
Calcitonin
Beta-lactoglobulin
2004
07
01
158
163
https://www.ijbiotech.com/article_6936_480e6518a88c715b47a4425199803d3a.pdf
Iranian Journal of Biotechnology
IRAN J BIOTECH
1728-3043
1728-3043
2004
2
3
Development of PCR-ELISA Technique for Determination of HLADRB1*01 Group Alleles
Habib
Nasiri
Mehdi
Forouzandeh
Aliakbar
Pourfathollah
Mohammad Javad
Rasaee
Fatemeh
Rahbarizadeh
We have developed an allotyping assay for detection of four HLA DRB1*01 group alleles based on polymerasechain reaction and solution hybridization in a microtiter plate. Using group specific primers a regionwithin exon 2 of HLA DRB1 gene was amplified by PCR. Labeling of PCR product was achieved byadding small amount of Dig-dUTP in place of dTTP. Labeled PCR product was hybridized to allele (HLADRB1*0101, *0102, *0103 and *0104) specific and a group (HLA DRB1*01) specific oligonucleotide probesin separate wells of the plate. Hybridized amplicones were detected by an enzymatic procedure. NinetyDNA samples were tested in parallel with PCR-SSP typing. The results were found to be well correlated bytwo methods. These results further suggest that, PCRELISA would be a rapid, specific and simple methodthat can be used for high resolution HLA typing before bone marrow and stem cell transplantation.
HLA DRB1
PCR-ELISA
Hybridization
Group specific amplification
2004
07
01
164
169
https://www.ijbiotech.com/article_6905_51cd755ebf5d6cc0573fefc000d8a51f.pdf
Iranian Journal of Biotechnology
IRAN J BIOTECH
1728-3043
1728-3043
2004
2
3
High-level expression and evaluation of the antigenicity of a recombinant Toxoplasma gondii GRA2 Protein
Majid
Golkar
Sima
Rafati
Yasaman
Taslimi
Tahereh
Taheri
Fatemeh
Doustdari
Mehdi
Assmar
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 Toxoplasmagondii, which induces strong antibody and T-cell response in human and mice. The purpose of thisstudy was to prepare recombinant GRA2 and evaluate its antigenic properties using infected mice sera. Toreach this point, GRA2 gene was highly expressed as a fusion protein with Thioredoxin (TRX) in Escherichiacoli BL21pLysS strain. The protein was purified in a single step on Ni-NTA affinity column. TRX-GRA2 wasconfirmed by Western blot technique using a monoclonal antibody specific for GRA2. Sera from miceinfected with Toxoplasma gondii showed high reactivity toward GRA2 and the level of IgG2a isotype waspredominant and significantly higher than IgG1.Taken together, TRX-GRA2 might be considered as an idealantigen to be used as a vaccine target as well as diagnostic tool for detection of toxoplasmosis.
Toxoplasma gondii
GRA2
Thioredoxin
E. coli
Expression
2004
07
01
170
176
https://www.ijbiotech.com/article_6907_157af281291b57b891b85b3d95a6f140.pdf
Iranian Journal of Biotechnology
IRAN J BIOTECH
1728-3043
1728-3043
2004
2
3
Disruption of GAL1 Gene in Saccharomyces cerevisiae Leads to Higher Ethanol Production
Abbas
Rezaee
Hyun Ah
Kang
Sang Ki
Rhee
GAL gene family is a set of structural and regulatory genes that enables cells to utilize galactose as a carbonsource in Saccharomyces cerevisiae. Phosphorylation of intracellular galactose can be catalyzed bygalactokinase (encoded by GAL1 gene). In this study role of GAL1 gene on ethanol production by S. cerevisiaetogether 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.
GAL1
Saccharomyces Cerevisiae
Gene disruption
Ethanol
2004
07
01
177
82
https://www.ijbiotech.com/article_6908_8ffa66790224dab6eb3d7b2061bef2b4.pdf
Iranian Journal of Biotechnology
IRAN J BIOTECH
1728-3043
1728-3043
2004
2
3
Cloning, expression and purification of Clostridium botulinum neurotoxin type E binding domain
Mir Latif
Mousavi
Shideh
Montaser Kouhsari
Shahram
Nazarian
Iraj
Rasooli
Jafar
Amani
Botulinum neurotoxins constitute a family of bacterial toxins for botulism syndrome in human. The toxinsbind with high affinity to nerve cells where they cause a complete inhibition and release of neurotransmittersand thereby produce flaccid paralysis. In this study the binding domain of type E neurotoxin was isolated byPCR and expressed in a proper expression vector. The results of this investigation can be used as a toolto study the mechanism of binding of holotoxins. This study is also implicated in antibody production againstbotulism syndrome.
Botulinum neurotoxin type E
Binding Domain
Expression
2004
07
01
183
188
https://www.ijbiotech.com/article_6909_79f3057bec43722c922ee415a34c6977.pdf
Iranian Journal of Biotechnology
IRAN J BIOTECH
1728-3043
1728-3043
2004
2
3
Purification of tyrosinase from edible mushroom
Kamahldin
Haghbeen
Ferdous
Rastgar Jazii
Ali Asghar
Karkhane
Shahrzad
Shareefi Borojerdi
A simple preparative method was developed for purification of Tyrosinase from edible mushroom (Agaricusbispora). 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 andchromatographed on a DE-52 column. On the basis of the activities assay, the eluted fractions by 150 mMsalt solution were selected for further purification. The collected fractions were pooled and chromatographedon a Sephadex G-200 column. Polyacrylamide gel electrophoresis (PAGE) of the purified tyrosinase produceda single band right beside the commercial sample obtained from Sigma Company at 128 kDa. Thelyophilized form of the purified Tyrosinase had a purification degree of 104 and showed strong cresolaseand catecholase activities when compared to a commerically available tyrosinase.
Tyrosinase
Edible mushroom
Purification
Extraction
2004
07
01
189
194
https://www.ijbiotech.com/article_6910_2ee60dd3ea3eee72653e5c43423e3bb7.pdf
Iranian Journal of Biotechnology
IRAN J BIOTECH
1728-3043
1728-3043
2004
2
3
Comparative analyses of the genetic diversity among bread wheat genotypes based on RAPD and SSR markers
Mohammad Reza
Naghavi
Mohsen
Mardi
Hossein Ali
Ramshini
Bahman
Fazelinasab
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 wheatcultivars/lines can be obtained by using RAPDs alone or in combination with SSRs, but SSRs alone cannotbe used for this purpose. Both techniques discriminated the genotypes very effectively. On the hand,RAPDs were able to discriminate the cultivars Alvand and Ghods, whereas the cultivars Sardari and Ghodswere discriminated only by SSRs. The use of PCRbased assays having advantage of being quick, easyto use and refractory to many environmental influences can complement traditional methods of germplasm characterization.
Wheat
Diversity
RAPD
SSR
2004
07
01
195
202
https://www.ijbiotech.com/article_6911_dcbb61f4f6658e6cf7e07f63f45aa71b.pdf
Iranian Journal of Biotechnology
IRAN J BIOTECH
1728-3043
1728-3043
2004
2
3
Detection of NAD(P)H: quinone oxidoreductase 609C T polymorphism in blood and archival human tissues using a simple PCR method
Firouzeh
Biramijamal
Mohammed Hossein
Sanati
Guity
Iravanloo
Kourosh
Shamimi
Dariush D.
Farhud
NAD(P)H: quinone oxidoreductase (NQO1) plays an important role in detoxification of numerous endogenousand foreign compounds. This gene has a single nucleotide polymorphism at site of codon 187(CCT TCT). Recently, it has been demonstrated that individuals with T allele may exhibit resistance toquinone based anticancer drugs such as mitomycin C. In the present study, a simple and feasible method wasdeveloped for detection of NQO1 genotype. In this modified procedure, dimethylsulfoxide (DMSO) andTriton X-100 were eliminated, also, PCR cycling conditions were modified to improve the PCR products fromblood and formalin-fixed, paraffin-embedded tissues. PCR-RFLP and DNA sequencing analysis carried outon a limited number of blood and archival samples. It is suggested that this procedure convenient for NQO1genotyping.
NQO1
Polymorphism
simple PCR-RFLP method
2004
07
01
203
206
https://www.ijbiotech.com/article_6912_8474984e4ad22a9ff1c3413f536381e3.pdf