Cell-free protein synthesis (CFPS) is an in vitro process that enables diverse applications in research, biomanufacturing, point-of-care diagnostics, therapeutics, and education using minimal laboratory equipment and reagents. One of the major limitations of CFPS implementation is its sensitivity to plasmid type. Specifically, plasmid templates based on commonly used vector backbones such as the pET series of bacterial expression vectors result in the inferior production of proteins. To overcome this limitation, we have evaluated the effect of expression cassette elements present in the pET30 vector on protein production across three different CFPS systems: NEBExpress, PURExpress, and CFAI-based E. coli extracts. Through the systematic elimination of genetic elements within the pET30 vector, we have identified elements that are responsible for the poor performance of pET30 vectors in the various CFPS systems. As a result, we demonstrate that through the removal of the lac operator (lacO) and N-terminal tags included in the vector backbone sequence, a pET vector can support high titers of protein expression when using extract-based CFPS systems. This work provides two key advances for the research community: 1) identification of vector sequence elements that affect robust production of proteins; 2) evaluation of expression across three unique CFPS systems including CFAI extracts, NEBexpress, and PURExpress. We anticipate that this work will improve access to CFPS by enabling researchers to choose the correct expression backbone within the context of their preferred expression system.
High-school and undergraduate college students may often rely on memorization when learning periodic trends, leading to an ineffective understanding of introductory chemistry concepts like atomic radius, ionization energy, and electron affinity. Comprehension of effective nuclear charge (Zeff) is foundational to a complete understanding of periodic trends. Zeff remains an abstract concept for many students, indicating that a manipulative activity for teaching the phenomenon could be a useful way to explain the concept for greater student comprehension. Here we report a hands-on learning activity to target this knowledge gap using magnetic attraction as an analogy for electrostatic attraction within the atom. This approach enables students to explore the phenomenon of effective nuclear charge at a macroscopic level and apply their learning to periodic trends and related concepts. We anticipate that this activity will fill a long-standing hole in the active learning of chemical principles. Importantly, this activity is low-cost and can be assembled using readily accessible materials to allow implementation in most classrooms and virtual learning environments.
High-school and undergraduate college students may often rely on memorization when learning periodic trends, leading to an ineffective understanding of introductory chemistry concepts like atomic radius, ionization energy, and electron affinity. Comprehension of effective nuclear charge (Zeff) is foundational to a complete understanding of periodic trends. Zeff remains an abstract concept for many students, indicating that a manipulative activity for teaching the phenomenon could be a useful way to explain the concept for greater student comprehension. Here we report a hands-on learning activity to target this knowledge gap using magnetic attraction as an analogy for electrostatic attraction within the atom. This approach enables students to explore the phenomenon of effective nuclear charge at a macroscopic level and apply their learning to periodic trends and related concepts. We anticipate that this activity will fill a long-standing hole in the active learning of chemical principles. Importantly, this activity is low-cost and can be assembled using readily accessible materials to allow implementation in most classrooms and virtual learning environments.
High-school and undergraduate college students may often rely on memorization when learning periodic trends, leading to an ineffective understanding of introductory chemistry concepts like atomic radius, ionization energy, and electron affinity. Comprehension of effective nuclear charge (Zeff) is foundational to a complete understanding of periodic trends. Zeff remains an abstract concept for many students, indicating that a manipulative activity for teaching the phenomenon could be a useful way to explain the concept for greater student comprehension. Here we report a hands-on learning activity to target this knowledge gap using magnetic attraction as an analogy for electrostatic attraction within the atom. This approach enables students to explore the phenomenon of effective nuclear charge at a macroscopic level and apply their learning to periodic trends and related concepts. We anticipate that this activity will fill a long-standing hole in the active learning of chemical principles. Importantly, this activity is low-cost and can be assembled using readily accessible materials to allow implementation in most classrooms and virtual learning environments.
High-school and undergraduate college students may often rely on memorization when learning periodic trends, leading to an ineffective understanding of introductory chemistry concepts like atomic radius, ionization energy, and electron affinity. Comprehension of effective nuclear charge (Zeff) is foundational to a complete understanding of periodic trends. Zeff remains an abstract concept for many students, indicating that a manipulative activity for teaching the phenomenon could be a useful way to explain the concept for greater student comprehension. Here we report a hands-on learning activity to target this knowledge gap using magnetic attraction as an analogy for electrostatic attraction within the atom. This approach enables students to explore the phenomenon of effective nuclear charge at a macroscopic level and apply their learning to periodic trends and related concepts. We anticipate that this activity will fill a long-standing hole in the active learning of chemical principles. Importantly, this activity is low-cost and can be assembled using readily accessible materials to allow implementation in most classrooms and virtual learning environments.
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