2 June 30, 2015
1. Grigory A. Gerashchenkov, Gulnar R. Yasybaeva, Nataliya A. Rozhnova, Aleksey V. Chemeris
Isolation of Promoters and Fragments of Genes Controlling Endosperm Development Without Fertilization in Arabidopsis and Engineering of the Antisense Constructions
European Journal of Molecular Biotechnology, 2015, Vol.(8), Is. 2, pp. 56-62.
Number of views: 2479 Download in PDF
2. Ignat Ignatov, Oleg MosinEuropean Journal of Molecular Biotechnology, 2015, Vol.(8), Is. 2, pp. 56-62.
Abstract:
Apomixis is asexual seed reproduction without both meiosis and fertilization based on the complex developmental processes such as apomeiosis, parthenogenesis and specific endosperm development. This investigation is aimed at engineering of apomixis in Arabidopsis thaliana with sexual seed reproduction. The fragments of known genes of endosperm formation MEA, FIE, FIS2 and gene of apomeiosis DYAD (as control) were isolated using Q5 high fidelity DNA polymerase. These gene fragments of interest at the antisense orientation were fused with isolated constitutive and meiosis specific promoters of Arabidopsis at NcoI sites. The fused promoter-gene fragment modules were cloned in pCambia1301 at SalI cites. The engineered constructions will be used for the floral dip transformation of Arabidopsis and down regulation of these genes at engineering of apomixis.
Apomixis is asexual seed reproduction without both meiosis and fertilization based on the complex developmental processes such as apomeiosis, parthenogenesis and specific endosperm development. This investigation is aimed at engineering of apomixis in Arabidopsis thaliana with sexual seed reproduction. The fragments of known genes of endosperm formation MEA, FIE, FIS2 and gene of apomeiosis DYAD (as control) were isolated using Q5 high fidelity DNA polymerase. These gene fragments of interest at the antisense orientation were fused with isolated constitutive and meiosis specific promoters of Arabidopsis at NcoI sites. The fused promoter-gene fragment modules were cloned in pCambia1301 at SalI cites. The engineered constructions will be used for the floral dip transformation of Arabidopsis and down regulation of these genes at engineering of apomixis.
Number of views: 2479 Download in PDF
Studying the Mechanism of Phototransformation of Light Signal by Various Mammal and Bacterial Photoreceptor Pigments - Rhodopsin, Iodopsin and Bacteriorhodopsin
European Journal of Molecular Biotechnology, 2015, Vol.(8), Is. 2, pp. 63-79.
Number of views: 2385 Download in PDF
3. Da´ad Ali HussainEuropean Journal of Molecular Biotechnology, 2015, Vol.(8), Is. 2, pp. 63-79.
Abstract:
This review article outlines the structure and function of mammal and bacterial photoreceptor pigments (rhodopsin, iodopsin, bacteriorhodopsin) and their aspects of bio-nanotechnological usage. On an example of bacteriorhodopsin is described the method of its isolation from purple membranes of photo-organotrophic halobacterium Halobacterium halobium ET 1001 by cellular autolysis by distilled water, processing of bacterial biomass by ultrasound at 22 KHz, alcohol extraction of low and high-weight molecular impurities, cellular RNA, carotenoids and lipids, the solubilization with 0,5 % (w/v) SDS-Na and subsequent fractionation by methanol and gel filtration chromatography on Sephadex G-200 Column balanced with 0,09 M Tris-buffer (pH = 8,35) with 0,1 % (w/v) SDS-Na and 2,5 mM EDTA. Within the framework of the research the mechanism of color perception by the visual retina analyzer having the ability to analyze certain ranges of the optical spectrum as colors, was studied along with an analysis of the additive mixing of two or more colors. It was shown that at the mixing of electromagnetic waves with different wavelengths, the visual analyzer perceives them as the separate or average wave length corresponding to the mixing color.
This review article outlines the structure and function of mammal and bacterial photoreceptor pigments (rhodopsin, iodopsin, bacteriorhodopsin) and their aspects of bio-nanotechnological usage. On an example of bacteriorhodopsin is described the method of its isolation from purple membranes of photo-organotrophic halobacterium Halobacterium halobium ET 1001 by cellular autolysis by distilled water, processing of bacterial biomass by ultrasound at 22 KHz, alcohol extraction of low and high-weight molecular impurities, cellular RNA, carotenoids and lipids, the solubilization with 0,5 % (w/v) SDS-Na and subsequent fractionation by methanol and gel filtration chromatography on Sephadex G-200 Column balanced with 0,09 M Tris-buffer (pH = 8,35) with 0,1 % (w/v) SDS-Na and 2,5 mM EDTA. Within the framework of the research the mechanism of color perception by the visual retina analyzer having the ability to analyze certain ranges of the optical spectrum as colors, was studied along with an analysis of the additive mixing of two or more colors. It was shown that at the mixing of electromagnetic waves with different wavelengths, the visual analyzer perceives them as the separate or average wave length corresponding to the mixing color.
Number of views: 2385 Download in PDF
Molecular Characterization of Some Productive Traits in Mesopotamian Buffaloes (Bubalus bubalis)
European Journal of Molecular Biotechnology, 2015, Vol.(8), Is. 2, pp. 80-87.
Number of views: 2361 Download in PDF
4. Oleg Mosin, Ignat IgnatovEuropean Journal of Molecular Biotechnology, 2015, Vol.(8), Is. 2, pp. 80-87.
Abstract:
One of these major genes is growth hormone and growth hormone receptor (GH, GHR) genes which are related to production and reproduction traits in livestock. Associations were analysed between the bovine growth hormone (GH) as well as growth hormone receptor (GHR) combined with their genotypes (GH/AluI& GHR/AluI). There is extensive literature on the genetic polymorphism of GH and GHR in cattle, but perusal of literature has indicated paucity of information on these two genes in buffalo. This study aimed to evaluate the genetic polymorphism within growth hormone and its receptor genes in Iraqi buffalo using PCR-RFLP technique. Genomic DNA extracted from 100 healthy buffaloes and amplified using primers that were designed from the cattle GH and GHR gene sequences. All tested buffalo DNA amplified fragments at 428bp for GH 5th exon. The digestion with AluI endonuclease gave three types of genotypes. The Distribution of the three genotypes and allele frequency was calculated according to Hardy-Weinberg equation, were LL=94 (94.09%), LV=6 (5.82%) and VV=0 (0.09%), so that most of buffaloes was homozygous (LL) due to the presence of the restriction site at position 52^53 (AG^CT) and less heterozygous was for the valine allele as compared with leucine allele homozygous. The amplified fragments of GHR gene obtained from all tested buffalo DNA at 342-bp were digested with AluI endonuclease. The result showed that all tested buffaloes are genotyped as GG for GHR gene, where amplified fragments were digested into two digested fragments at 241and 101-bp due to the presence of AluI restriction site at position (AG^CT).
One of these major genes is growth hormone and growth hormone receptor (GH, GHR) genes which are related to production and reproduction traits in livestock. Associations were analysed between the bovine growth hormone (GH) as well as growth hormone receptor (GHR) combined with their genotypes (GH/AluI& GHR/AluI). There is extensive literature on the genetic polymorphism of GH and GHR in cattle, but perusal of literature has indicated paucity of information on these two genes in buffalo. This study aimed to evaluate the genetic polymorphism within growth hormone and its receptor genes in Iraqi buffalo using PCR-RFLP technique. Genomic DNA extracted from 100 healthy buffaloes and amplified using primers that were designed from the cattle GH and GHR gene sequences. All tested buffalo DNA amplified fragments at 428bp for GH 5th exon. The digestion with AluI endonuclease gave three types of genotypes. The Distribution of the three genotypes and allele frequency was calculated according to Hardy-Weinberg equation, were LL=94 (94.09%), LV=6 (5.82%) and VV=0 (0.09%), so that most of buffaloes was homozygous (LL) due to the presence of the restriction site at position 52^53 (AG^CT) and less heterozygous was for the valine allele as compared with leucine allele homozygous. The amplified fragments of GHR gene obtained from all tested buffalo DNA at 342-bp were digested with AluI endonuclease. The result showed that all tested buffaloes are genotyped as GG for GHR gene, where amplified fragments were digested into two digested fragments at 241and 101-bp due to the presence of AluI restriction site at position (AG^CT).
Number of views: 2361 Download in PDF
Metabolism, Physiology and Biotechnological Applications of Halobacteria
European Journal of Molecular Biotechnology, 2015, Vol.(8), Is. 2, pp. 88-102.
Number of views: 2514 Download in PDF
5. Sajedeh Saeedfar, Marzieh Negahban, Mohammad Mahmoodi SoorestaniEuropean Journal of Molecular Biotechnology, 2015, Vol.(8), Is. 2, pp. 88-102.
Abstract:
Halophiles (lat. “salt-loving”) is the taxonomic group of extreme aerobic obligate Gram-negative microorganisms that live in conditions of high salinity – in the seas, salt lakes, saline soils etc. These microorganisms are known to reddish patina on products, preserved with using large quantities of salt (NaCl). Halophiles were isolated for the first time at the beginning of the XX century from the marine flora estuary mud, but their systematic study was started only at the end of the second decade of the XX century. The internal environment of the human body is not suitable for existence of halobacteria, since none of them are known to have pathogenic forms. Halobacteria have great practical potential for using in molecular bioelectronics and bio-nanotechnology due to their unique ability to convert the energy of sunlight into electrochemical energy of protons H+ due to the presence in their cells a special photo transforming retinal containing integral protein – bacteriorhodopsin, the mechanism of action of which has been currently studied in detail. This article describes the characteristics of the metabolism and physiology of halophilic bacteria, as well as a method of biosynthesis and preparation of bacteriorhodopsin from purple membranes of cells of the extreme photoorganotrophic halobacterium Halobacterium halobium.
Halophiles (lat. “salt-loving”) is the taxonomic group of extreme aerobic obligate Gram-negative microorganisms that live in conditions of high salinity – in the seas, salt lakes, saline soils etc. These microorganisms are known to reddish patina on products, preserved with using large quantities of salt (NaCl). Halophiles were isolated for the first time at the beginning of the XX century from the marine flora estuary mud, but their systematic study was started only at the end of the second decade of the XX century. The internal environment of the human body is not suitable for existence of halobacteria, since none of them are known to have pathogenic forms. Halobacteria have great practical potential for using in molecular bioelectronics and bio-nanotechnology due to their unique ability to convert the energy of sunlight into electrochemical energy of protons H+ due to the presence in their cells a special photo transforming retinal containing integral protein – bacteriorhodopsin, the mechanism of action of which has been currently studied in detail. This article describes the characteristics of the metabolism and physiology of halophilic bacteria, as well as a method of biosynthesis and preparation of bacteriorhodopsin from purple membranes of cells of the extreme photoorganotrophic halobacterium Halobacterium halobium.
Number of views: 2514 Download in PDF
The Effect of Drought Stress on the Essential Oil Content and Some of the Biochemical Characteristics of Anise Hyssop (Agastache foeniculum [Pursh] Kuntze)
European Journal of Molecular Biotechnology, 2015, Vol.(8), Is. 2, pp. 103-114.
Number of views: 3066 Download in PDF
6. European Journal of Molecular Biotechnology, 2015, Vol.(8), Is. 2, pp. 103-114.
Abstract:
Plot trials were carried out in a research field in Tehran (Iran) to determine the effect of drought stress on the essential oil content and some of the plant biochemical characteristics of Anise Hyssop (Agastache foeniculum [Pursh] Kuntze), which is a valuable medicinal plant. Drought stress was conducted at different levels including: well-water (100% FC), mild drought stress (85% FC), moderate drought stress (70% FC), severe drought stress (55% FC), 100% FC (vegetative stage) 85% FC (reproductive stage), 100% FC (vegetative stage) 70% FC (reproductive stage), and 85% FC (vegetative stage) 100% FC (reproductive stage). The experiment was arranged as a RCBD with three replications. The output results showed that, water deficit stress significantly (P≤0.05) increased activities of antioxidant enzymes (Superoxide Dismutase, Catalase, Glutathione Peroxidase) as well as Essential Oil yield and Abscisic Acid content. Lipid and protein oxidation (malondialdehyde and dityrosine contents) also increased significantly under severe water deficit stress. According to the results, severe drought conduction (55% FC) is the optimum level of soil moisture to plant Anise Hyssop under water deficit stress.
Plot trials were carried out in a research field in Tehran (Iran) to determine the effect of drought stress on the essential oil content and some of the plant biochemical characteristics of Anise Hyssop (Agastache foeniculum [Pursh] Kuntze), which is a valuable medicinal plant. Drought stress was conducted at different levels including: well-water (100% FC), mild drought stress (85% FC), moderate drought stress (70% FC), severe drought stress (55% FC), 100% FC (vegetative stage) 85% FC (reproductive stage), 100% FC (vegetative stage) 70% FC (reproductive stage), and 85% FC (vegetative stage) 100% FC (reproductive stage). The experiment was arranged as a RCBD with three replications. The output results showed that, water deficit stress significantly (P≤0.05) increased activities of antioxidant enzymes (Superoxide Dismutase, Catalase, Glutathione Peroxidase) as well as Essential Oil yield and Abscisic Acid content. Lipid and protein oxidation (malondialdehyde and dityrosine contents) also increased significantly under severe water deficit stress. According to the results, severe drought conduction (55% FC) is the optimum level of soil moisture to plant Anise Hyssop under water deficit stress.
Number of views: 3066 Download in PDF