PCR assembly of synthetic human erythropoietin gene

Human erythropoietin (huEPO) is a glycoprotein with important physiological functions, such as erythropoiesis, angiogenesis, and wound healing. A therapeutic protein, huEPO is commonly used to treat patients suffering from renal and non-renal anemia. Recombinant human erythropoietin (rhuEPO) and endogenous huEPO are similar with respect to their biological and chemical properties. In this study, we describe the construction of synthetic huEPO gene to produce rhuEPO. The synthetic huEPO gene was constructed by overlapping oligonucleotides assembly and amplified by polymerase chain reaction (PCR). Twenty oligonucleotide sets, covering the huEPO gene sequence and two newly introduced restriction enzyme sites, were pulled together and amplified using Pfu DNA polymerase to produce the expected DNA products with sizes of ~500bp and ~600bp. The PCR products were ligated into pGEM-T plasmid vector to facilitate DNA sequencing process of the constructed huEPO gene and downstream cloning manipulation. DNA sequence analysis showed correctly assembled oligonucleotide sets, representing the huEPO gene sequence albeit with minor base mutations. Hence, oligonucleotides assembly and PCR amplification provide a convenient and speedy method for the synthesis of huEPO gene without depending on mRNA isolation and reverse transcription or the need to have a genomic library

Green potential rating tool: an assessment of green potential for conventional buildings

Assessment of the green potentials in a conventional building is rarely discussed in past literature unlike other types of assessments, such as a building’s current performance and qualities. ‘Green potential’ is the capacity of a conventional building to be refurbished into a green building. This paper presents the development of a rating tool to assess green potentials of existing conventional buildings. The development process involves reviewing relevant literature on the existing assessment tools. The review focuses on identifying methods and indicators that can be adopted for the assessment of green potentials. It is discovered that while literature on green potential assessment is limited, the frameworks of other types of assessments concerning green buildings are still suitable to be adopted. Additionally, with some modifications, commercial green building rating tools provide the most suitable indicators to assess green potentials. Apart from filling the knowledge gap, the tool developed may also assist building managers strategize towards achieving sustainability for large building stocks such as a small township or a university campus

Case note 2013 (5748)

Injection molding of fiber-reinforced polymer composites is associated with the problem of fiber breakage. If the fiber length retained in the finish product is too short, it will limit the expected improvement in the property. Extrusion and pultrusion are two methods normally employed for the melt compounding of polymer composite feedstock for the injection molding and produced short- and long-fiber composites (SFCs and LFCs), respectively. In this work, short-and long-glass fiber-reinforced polyamide 6,6 composites were injection molded at different fiber loading and tested for the tensile properties, impact properties and fiber length characteristics. It was found that both tensile strength and tensile modulus of LFCs improved compared to the SFCs counterpart despite reduction in fracture strain, while pultrusion compounded composites also showed superior fiber characteristics, in terms of fiber length distribution, L n, L w, etc. compared to the extrusion compounded composites counterpart. Fiber length characteristics were also in agreement with the improvement in tensile strength and tensile modulus of LFCs over the SFCs. Impact properties of LFCs also show some improvement compared to the SFCs counterpart with equivalent composition, despite longer fiber retained in composites

PCR assembly of synthetic human erythropoietin gene

Human erythropoietin (huEPO) is a glycoprotein with important physiological functions, such as erythropoiesis, angiogenesis, and wound healing. A therapeutic protein, huEPO is commonly used to treat patients suffering from renal and non-renal anemia. Recombinant human erythropoietin (rhuEPO) and endogenous huEPO are similar with respect to their biological and chemical properties. In this study, we describe the construction of synthetic huEPO gene to produce rhuEPO. The synthetic huEPO gene was constructed by overlapping oligonucleotides assembly and amplified by polymerase chain reaction (PCR). Twenty oligonucleotide sets, covering the huEPO gene sequence and two newly introduced restriction enzyme sites, were pulled together and amplified using Pfu DNA polymerase to produce the expected DNA products with sizes of ~500bp and ~600bp. The PCR products were ligated into pGEM-T plasmid vector to facilitate DNA sequencing process of the constructed huEPO gene and downstream cloning manipulation. DNA sequence analysis showed correctly assembled oligonucleotide sets, representing the huEPO gene sequence albeit with minor base mutations. Hence, oligonucleotides assembly and PCR amplification provide a convenient and speedy method for the synthesis of huEPO gene without depending on mRNA isolation and reverse transcription or the need to have a genomic library