Production of porcine cloned transgenic embryos expressing green fluorescent protein by somatic cell nuclear transfer

Abstract

The increases of H2O2 concentrations in plant cells often occur under biotic and abiotic stress conditions (e.g. light, environmental stresses and plant hormone abscisic acid). Atmospheric H202 as an ancient signal molecule not only plays the key role in inducing evolution of oxygenic photosynthesis, but also modulates many physiological events, such as stomatal movement, hypersensitive responses, programmed cell death and gene expressions. H2O2 levels in cells must sustain a fine equilibrium between production and scavenging. H2O2 enters cells from the apoplast or generated sources, and in turn is distributed in sub-cellular compartments. H202 can modulate the activities of many components in signaling, such as protein phosphatases, protein kinases, transcription factors (TFs), and calcium channels. Elevated cytosolic calcium concentrations will initiate further downstream responses, via the action of calcium-binding proteins. On the other hand, the research of H2O2 as a signal molecule is still in a comparatively juvenile stage, for example, little is known about how the cells sense H2O2, what the rate-limiting steps and most important cellular events are in cell signaling and what kind of genes is specific or necessary to H2O2 signaling. The answers to all the questions depend on the functional genomic and molecular genetics analysis.

Overexpression of Arachis hypogaea AREB1 Gene Enhances Drought Tolerance by Modulating ROS Scavenging and Maintaining Endogenous ABA Content

“…For H 2 O 2 detection, the treated samples were immersed in 1 mg/mL fresh DAB solution (pH 5.8) (prepared in 10 mmol/L phosphate buffer, pH 3.8) and incubated in dark at 28 °C for 8 h. The stained samples were then bleached in 80% ethanol and boiled for several minutes (2–5 min), then dropped for staining by concentrated ethanol and kept in 4 °C, as described by Cheng and Song [50]. …”

Construction of Recombinant Marek's Disease Virus (rMDV) Co-Expressing AIV-H9N2-NA and NDV-F Genes under Control of MDV's Own Bi-Directional Promoter

“…More in vitro studies from our laboratory demonstrated that the promoter could drive the expression of GFP or CAT as reporter genes in each direction with the presence of MDV pp24/pp38 hetero-dimer. The pp24/38 dimer was able to bind to the promoter and worked in-trans as a transcriptional factor to enhance the transcription activity [9], [41]. In this study, use recombinant plasmid transient transfected CEF as well as in recombinant MDV infected CEF, it was demonstrated that different foreign genes could be expressed simultaneously under control of the bi-directional promoter.…”

“…The results clearly indicated that the MDV endogenous bi-directional promoter of only 305 bp could effectively drive two foreign genes expressed in two directions. As mentioned above, the bi-directional promoter is located between pp38 and 1.8kb mRNA family transcription start sites and pp38/pp24 dimer could work as trans-acting transcriptional factor to strongly enhance its promoter activity [8], [9], [41], the bi-directional promoter may have some advantages in expression of two foreign genes in the early stage of MDV infection. This is because pp38 is an early gene related to MDV early B-lymphocyte cytolytic infection [44], and pp38/pp24 dimers should be available in the early stage of infection.…”

“…The 1.8-kb mRNA transcript family [3], [4] and the 38kd phosphorylated protein gene (pp38) [5], [6] are two of many MDV-specific genes. They were located on I RL region and separated by a short sequence of only about 305 bp but with several enhancer motifs such as TATA-box, CAAT-box, Oct-1, and Sp1, and as a bi-directional promoter to initiate transcription of two genes in opposite directions [7], [8], [9]. By use of GFP gene and chloramphenicol acetyltransferase ( CAT ) as reporter genes, it was demonstrated that the activity of the bi-directional promoter could be strongly increased by the expression of MDV pp38/pp24 dimers as a trans-acting transcriptional factor [10].…”

Biotechnological bases of the development of cloned pig embryos

“…The matured oocytes at the stage of metaphase II with the visualized first polar body are usually used as sources of cytoplasts for cloning (Hardarson et al, 2000). In pigs, the optimal duration of in vitro maturation of oocytes varies according to different studies in the range from 24 to 44 hours (Zhang et al, 2006;Sugimura et al, 2010).…”