eng
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2003-10-01
1
4
197
206
6902
Enzyme Immobilization: The State of Art in Biotechnology
Dariush Norouzian
dnsa@pasteur.ac.ir
1
Pilot Biotechnology Department, Pasteur Institute of Iran,Tehran, I.R. Iran.
The advantages of immobilized enzyme over its soluble counterpart arise from their improved stability andeasy separation from the reaction media, leading to decrease in production cost. Immobilization methodsrange from adsorption onto matrices, entrapment, cross-linking and covalent bonding to prefabricatedcarriers or activated supports. Changes in kinetic properties of immobilized enzyme can produce substrateor pH gradient, which reduce the reaction rates and finally product yields.
https://www.ijbiotech.com/article_6902_670b2ac4af9c3be208f1ffcd85ad7a06.pdf
Enzyme
Substrate
Matrix
Immobilization
eng
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2003-10-01
1
4
207
212
6884
Purification of Large Quantities of Biologically Active Recombinant Human Growth Hormone
Mahvash Khodabandeh
saba@nigep.ac.ir
1
Bagher Yakhchali
bahar@nigeb.ac.ir
2
Maryam Rahimi
3
Neda Vaseli
4
Zahra Moghaddasi Jahromi
5
Alireza Zomorrodipour
zomorodi1@gmail.com
6
Abdolkhalegh Deezagi
7
Saeid Ansari Majd
8
Mohammad H Sanati
9
National Research Center for Genetic Engineering and Biotechnology, P.O. Box: 14155-6343, Tehran, I.R. Iran.
National Research Center for Genetic Engineering and Biotechnology, P.O. Box: 14155-6343, Tehran, I.R. Iran.
National Research Center for Genetic Engineering and Biotechnology, P.O. Box: 14155-6343, Tehran, I.R. Iran.
National Research Center for Genetic Engineering and Biotechnology, P.O. Box: 14155-6343, Tehran, I.R. Iran.
National Research Center for Genetic Engineering and Biotechnology, P.O. Box: 14155-6343, Tehran, I.R. Iran.
National Research Center for Genetic Engineering and Biotechnology, P.O. Box: 14155-6343, Tehran, I.R. Iran.
National Research Center for Genetic Engineering and Biotechnology, P.O. Box: 14155-6343, Tehran, I.R. Iran.
National Research Center for Genetic Engineering and Biotechnology, P.O. Box: 14155-6343, Tehran, I.R. Iran.
National Research Center for Genetic Engineering and Biotechnology, P.O. Box: 14155-6343, Tehran, I.R. Iran.
Production and purification of human growth hormone using a simple method was studied in two recombinantEscherichia coli, D7-5 and C27-2 strains. The r-hGH was expressed in the form of inclusion body in a batchfermentation process and purified to 99% purity using a procedure based on acid precipitation of the hostderived proteins and other impurities. The effect of the pH and host strain on purification of the r-hGH andefficiency of the procedure were evaluated. It was found that the optimum pH for precipitation of the hostderived proteins was 4.9. The procedure was suitable for r-hGH purification from D7-5 stain but not from theother strain C27-2. The purity of > 99% and recovery of about 40% were obtained as shown by SDS-PAGEand Western blot analysis. The purified r-hGH was biologically active as judged by receptor assay withvery low endotoxin content which could be suitable for therapeutic applications. This simple and cost effectiveproduction process could be useful for large scale production of recombinant hGH from specific strains.
https://www.ijbiotech.com/article_6884_ae530adfc7eafeaf6f8669769d29841c.pdf
Recombinant protein
hGH
E. coli
Purification
eng
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2003-10-01
1
4
213
217
6880
Induction of Spawning in Common carp Cyprinus carpio,Using Pituitary Extract and GnRH Analogue in Combination with Domperidone
Salar Dorafshan
sdorafshan@gmail.com
1
Hossein Mostafavi
2
Bagher Mojazi Amiri
3
Department of Fisheries, College of Natural Resources, Isfahan University of Technology (IUT), Zip code:84154, Isfahan.
National Research Center of Genetic Engineering and Biotechnology (NRCGEB), P.O. Box:14155-6343, Tehran.
Department of Fisheries and Environmental Science, Faculty of Natural Resources, University of Tehran, P.O. Box: 31585-4314, Karaj, I.R. Iran.
The effectiveness of the first Iranian made gonadotropin releasing hormone analogue, [D-Ala6des-Gly10] GnRH ethylamide, alone or in combination with domperidone, a dopamine antagonist on spawningrate, latency period, working fecundity and embryo viability in common carp, Cyprinus carpio, was investigated.Fifty two fish were divided into 5 groups and treated intrapretoneally as follows: 3 mg/Kg b.w. of carp pituitary extract (C.P.E.) as a positive control, GnRHa alone, 10 μg/Kg b.w.. or in combination with domperidone, 5 mg/Kg b.w. in a single or double injections 7h apart. A group was treated with propylene glycol 0.2 ml/Kg b.w. alone and considered as control. No female ovulated in groups receiving either propylene glycol or 10 μg/Kg b.w. of GnRHa alone. The spawning rate was higher in female GnRHa+domperidone (10 μg/Kg b.w..+5 mg/Kg) in double injections (11 out of 12) as compared to fish which injected either with C.P.E (7 out of 16) or GnRHa + domperidone in a single injection (3 out of 12)(P<0.05). The mean working fecundity, was significantly higher for fish receiving GnRHa+domperidone in single (126214 ±24315) or double injections (145600 ± 27113) compared to C.P.E treated group (52435 ± 1224) (P<0.05). There were no significant differences for latency period or embryo viability among the groups.
https://www.ijbiotech.com/article_6880_8f9865a65e9104d21b343a9f0c01b38a.pdf
spawning induction
carp pituitary extract
GnRH analogue
domperidone
eng
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2003-10-01
1
4
218
223
6885
Initiation of Ageing Process by Meiotic and Mitotic Recombination within the Ribosomal DNA Genes in Saccharomyces cerevisiae
Majid Motovali-Bashi
mbashi@sci.ui.ac.ir
1
Zohreh Hojati
z.hojati@sci.ui.ac.ir
2
Richard Walmsley
3
Genetics group, Biology Department, Faculty of Sciences, Isfahan University, Isfahan, I. R. Iran.
Genetics group, Biology Department, Faculty of Sciences, Isfahan University, Isfahan, I. R. Iran.
Biomolecular Sciences Department, UMIST, Sackvill St., P.O. Box: 88, Manchester, M60 1QD, UK.
In the budding yeast of Saccharomyces cerevisiae the tandem repeated of rDNA genes are located onchromosome XII, which is in the nucleolus. There are different types of proteins in the nucleoluskeleton,silencing proteins have got important role in nucleolus.It is shown that meiotic recombination between nonsister chromatids in the rDNA genes are stronglysuppressed, and suggested that silencing proteins such as SIR2 are involved in silencing state. It is alsoshown that nucleolus shows some changes during ageing process. It is claimed that intrachromosomalrecombination within the rDNA repeated sequences; producing 3-μm rDNA circles are accumulated with oldcell. This study looked at the rDNA breakage in two different types of strain ORD 1181 according to theirage. The fine analysis of the rDNA array was performed using restriction endonuclease enzymes, whichdo not cleave within the rDNA array. The results suggest that there are only meiotic hot regions for chromosome breakage in the old cells within the rDNA array.
https://www.ijbiotech.com/article_6885_c7ddadb3eaba51539881a2866876a93b.pdf
Double-strand break (DSB)
rDNA gene
Meiotic recombination
ageing
(rDNA) homolog pairing
eng
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2003-10-01
1
4
224
233
6887
Sequence-Based Differentiation of Strains in the Streptomyces cyaneus Species-Group
Ehsan Rashidian
1
Michael Goodfellow
michael.goodfellow@ncl.ac.uk
2
Department of Microbiology, School of Veterinary Medicine, Lorestan University, Khorramabad, I.R. Iran.
2School of Biology, University of Newcastle, Newcastle Upon Tyne, NE1 7RU, United Kingdom.
In an extensive numerical phenetic survey, a numberof blue, red and gray spored streptomycete strainswere grouped together as Streptomyces cyaneusspecies-group, a taxon which encompasses strainsknown to produce antitumor antibiotics, notably anthracyclines.In the present investigation these and relatedstreptomycetes were the subject of morphological and16S rRNA sequencing studies designed to clarify theirtaxonomic relationships. It is evident from these resultsthat the Streptomyces cyaneus species-group encompassesmisclassified strains and members of severaldistinct species. However, some of the red-sporedstrains (e.g. Streptomyces janthinus ISP 5206T,Streptomyces roseoviolaceus ISP 5277T andStreptomyces violatus ISP 5209T) formed a distinctclade. In contrast, most of the blue and gray-sporedstrains showed much less sequence homologybetween and with one another and were scatteredthroughout the 16S rRNA Streptomyces tree. Theimproved classification of this group of streptomycetesprovides an essential basis for establishing their industrialand economical significance.
https://www.ijbiotech.com/article_6887_404ed6f454feb1f248afab7ae004c2a7.pdf
16S rDNA sequencing
Streptomycetes
Streptomyces cyaneus
eng
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2003-10-01
1
4
239
246
6881
A Heterologous Enzyme Linked Immunosorbant Assay of Morphine Using Penicillinase as Label
Shima Hallaj
1
Mohammad J. Rasaee
2
Monirsadat Haerian
3
Malihe Paknejad
4
Soheila Kashanian
kashanian_s@yahoo.com
5
Fatemeh Rahbarizadeh
rahbarif@modares.ac.ir
6
Kobra Omidfar
omidfar@tums.ac.ir
7
Mohammad Malekaneh
8
Mohammad Kakhki
9
Food and Drug Control Laboratories, Department of Biology, Tehran, Iran.
Department of Biochemistry, Faculty of Medical Sciences, Tarbiat Modarres University, Tehran, Iran.
Food and Drug Control Laboratories, Department of Biology, Tehran, Iran.
Department of Biochemistry, Faculty of Medical Sciences, Tarbiat Modarres University, Tehran, Iran.
Department of Biochemistry, Faculty of Medical Sciences, Tarbiat Modarres University, Tehran, Iran.
Food and Drug Control Laboratories, Department of Biology, Tehran, Iran.
Department of Biochemistry, Faculty of Medical Sciences, Tarbiat Modarres University, Tehran, Iran.
Department of Clinical Biochemistry, Birjand University of Medical Sciences, Birjand, Iran.
4Department of Pilot, Pasture Institute of Iran, Tehran, I.R. Iran.
A rapid, sensitive, specific and high through-put enzyme-linked immunosorbant assay (ELISA) methodfor determination of morphine in urine samples using penicillinase as label enzyme has been developed. Noextraction or chromatography was included in this assay procedure. Immunoglobulin (Ig) purified polyclonalanti-bodies against a C6-hemisuccinate derivative of morphine (M-C6-HS) conjugated to bovineserum albumin (BSA) was coated onto the wells of microtiter plate. A morphine-C3-hemisuccinate (M-C3-HS) was also prepared and the two derivatives were conjugated to penicillinase (M-C6-HS-P and M-C3-HSP).The heterologous combination of antibody prepared against M-C6-HS-BSA and enzyme conjugateprepared for M-C3-HS-P showed better properties in term of sensitivity, reproducibility and slope of standardcurve. The assay was sensitive from 20 pg/ml and detected up to 100 ng/ml of morphine in urine samples.The affinity of antibody in homologous assay was found to be 6.6×1010 l/mol and for heterologous assaywas 3.2 ×1012 l/mol. The assay was completed within 4 h. The homologous assays performed under differentconditions of coating, concentrations, duration, pH, etc. did not end up with a suitable standard curve.Hence it seems that the ability of morphine to displace the hapten enzyme conjugate dependds on the position of the enzyme coupled to the hapten molecule.This ELISA techniqu showed 100% correlation withimmunochromatography (IC) and 90% percent correlation with latex agglutination inhibition (LAI) test in theresults obtain with urine samples declared positive by authorities. ELISA also showed approximately 90%correlation with LAI-negative urine samples.
https://www.ijbiotech.com/article_6881_65b43fdb345e07a3c69bdefe71bb8cfd.pdf
ELISA
Morphine
Penicillinase
Heterology
eng
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2003-10-01
1
4
247
251
6886
Growth and Isolation of Human Cytomegalovirus on a New Human Fetal Foreskin Fibroblast-derived Cell Line in Iran
Samad Amini Bavil Olyaee
1
Farzaneh Sabahi
2
Mohammad Hassan Rostaie
3
Mohsen Karimi Arzenani
4
Zahra Samadi Bahrami
5
Ramin Sarrami Forooshani
6
Ahmad Adeli
7
Virology Department, School of medical Sciences, Tarbiat Modarres University, P.O. Box: 14115-111, Tehran, Iran.
Virology Department, School of medical Sciences, Tarbiat Modarres University, P.O. Box: 14115-111, Tehran, Iran.
Virology Department, School of medical Sciences, Tarbiat Modarres University, P.O. Box: 14115-111, Tehran, Iran.
Biotechnology Department, Pasteur Institute of Iran, Tehran, Iran.
3National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, I.R. Iran.
Biotechnology Department, Pasteur Institute of Iran, Tehran,l Iran.
Biotechnology Department, Pasteur Institute of Iran, Tehran, Iran.
Cell culture technique has been used for detection and confirmation of many different viruses in clinical samples. Although, new diagnostic methods have been developed for viral infections, traditional cell culturetechnique is still regarded as the “gold standard” for several infectious agents such as human cytomegalovirus(HCMV). In the present study, a new human fetal foreskin fibroblast (HFFF)-derived cell line was obtained from a normal Iranian male fetus at the end of first trimester at National Cell Bank of Iran (NCBI) and its ability for HCMV growth was investigated.Thirteen treated urine samples from renal transplant recipients were inoculated onto HFFF monolayer.HCMV growth was monitored and confirmed by typical CMV cytopathic effect (CPE), histological staining,immunological staining and polymerase chain reaction (PCR)-based restriction fragment length polymorphism(RFLP) method and sequencing. All of the abovementioneddetection methods were confirmed that newHFFF-derived cell line is a sensitive line for growth ofHCMV. This cell line has been named HFFF-PI6 and isavailable with accession number NCBI C170 at NCBIof Pasteur Institute of Iran.
https://www.ijbiotech.com/article_6886_da2091bd62559c436b9f33460efbd7ed.pdf
Human fetal foreskin fibroblast (HFFF)