ProxiMAX Randomisation:a new technology for non-degenerate saturation mutagenesis of contiguous codons

ProxiMAX randomisation achieves saturation mutagenesis of contiguous codons without degeneracy or bias. Offering an alternative to trinucleotide phosphoramidite chemistry, it uses nothing more sophisticated than unmodified oligonucleotides and standard molecular biology reagents and as such, requires no specialised chemistry, reagents nor equipment. When particular residues are known to affect protein activity/specificity, their combinatorial replacement with all 20 amino acids, or a subset thereof, can provide a rapid route to generating proteins with desirable characteristics. Conventionally, saturation mutagenesis replaced key codons with degenerate ones. Although simple to perform, that procedure resulted in unnecessarily large libraries, termination codons and inherent uneven amino acid representation. ProxiMAX randomisation is an enzyme-based technique that can encode unbiased representation of all or selected amino acids or else can provide required codons in pre-defined ratios. Each saturated position can be defined independently of the others. ProxiMAX randomisation is achieved via saturation cycling: an iterative process comprising blunt end ligation, amplification and digestion with a Type IIS restriction enzyme. We demonstrate both unbiased saturation of a short 6-mer peptide and saturation of a hypervariable region of a scfv antibody fragment, where 11 contiguous codons are saturated with selected codons, in pre-defined ratios. As such, ProxiMAX randomisation is particularly relevant to antibody engineering. The development of ProxiMAX randomisation from concept to reality is described.

ProxiMAX randomisation:a new technology for non-degenerate saturation mutagenesis of contiguous codons

Back in 2003, we published ‘MAX’ randomisation, a process of non-degenerate saturation mutagenesis using exactly 20 codons (one for each amino acid) or else any required subset of those 20 codons. ‘MAX’ randomisation saturates codons located in isolated positions within a protein, as might be required in enzyme engineering, or else on one face of an alpha-helix, as in zinc finger engineering. Since that time, we have been asked for an equivalent process that can saturate multiple, contiguous codons in a non-degenerate manner. We have now developed ‘ProxiMAX’ randomisation, which does just that: generating DNA cassettes for saturation mutagenesis without degeneracy or bias. Offering an alternative to trinucleotide phosphoramidite chemistry, ProxiMAX randomisation uses nothing more sophisticated than unmodified oligonucleotides and standard molecular biology reagents. Thus it requires no specialised chemistry, reagents nor equipment and simply relies on a process of saturation cycling comprising ligation, amplification and digestion for each cycle. The process can encode both unbiased representation of selected amino acids or else encode them in pre-defined ratios. Each saturated position can be defined independently of the others. We demonstrate accurate saturation of up to 11 contiguous codons. As such, ProxiMAX randomisation is particularly relevant to antibody engineering.

Founders' human capital and the performance of UK new technology based firms

This paper investigates the role of entrepreneurs' general and specific human capital on the performance of UK new technology based firms using a resource based approach to the entrepreneurship theory. The effect of entrepreneurial human capital on the performance of NTBFs is investigated using data derived from a survey of 412 firms operating in both high-tech manufacturing and the services sectors. According to the resource based theory it is found that specific human capital is more important for the performance of NTBFs in relation to general. More specifically individual entrepreneurs or entrepreneurial teams with high levels of formal business education, commercial, managerial or same sector experience are found to have created better performing NTBFs. Finally it is found that the performance of a NTBF can improve through the combination of heterogeneous but complementary skills, including, for example, technical education and commercial experience or managerial technical and managerial commercial experience. © 2010 Springer Science+Business Media, LLC.

Re-written by machine and new technology: Did the Internet kill the Video Star

The traditional way of understanding television content consumption and viewer reactions may be simply summarised: information about the program, viewing at airing time, and interpersonal discussion after the program. In our digital media environment due to crossmedia consumption and platform shifts, the actual trend in audiovisual, and traditionally television content consumption is changing, the viewer’s journey is different across contents and platforms. Content is becoming independent from the platform and the medium is increasingly in the hands of technologically empowered viewers. Our objective is to uncover how traditional content expressly manufactured for television (series, reality shows, sports) can now be consumed via other platforms, and how and to what extent audiovisual content consumption is complemented or replaced by other forms (text, audio). In our exploratory research we identify the typical patterns of interaction and synergies of consumption across classical media content. In this study we used a multimethodology qualitative research design with three research phases including focus groups, online content analysis, and viewers’ narratives. Overall, the Video Star stays alive, but has to deal with immediate reactions and has to change according to his or her audiences’ wishes

Factors Affecting the Adoption of New Technology: the case of 311 Government Call Centers

Government call centers (311) were first created to reduce the volume of non-emergency calls that were being placed to emergency 911 call centers. The number of 311 call centers increased from 57 in 2008 to about 300 in 2013. Considering that there are over 2,700 municipal government units across the United States, the adoption rate of the 311 centers is arguably low in the country. This dissertation is an examination of the adoption of 311 call centers by municipal governments. My focus is specifically on why municipal governments adopt 311 and identifying which barriers result in the non-adoption of 311 call centers. This dissertation is possibly the first study to examine the adoption of 311 call centers in the United States. The dissertation study has identified several significant factors in the adoption and non-adoption of 311 government call centers. The following factors were significant in the adoption of 311 government call centers: managerial support, financial constraints, organizational responsiveness, strategic plan placement, and technology champion. The following factors were significant barriers that resulted in the non-adoption of a 311 government call center; no demand from citizens, start up costs, annual operating costs, unavailability of funding, and no obvious need for one.If local government entities that do not have a 311 government call center decide to adopt one, this study will help them identify the conditions that need to be in place for successful adoption to occur. Local government officials would first need to address the barriers in setting up the 311 call centers.

Knowledge sources, innovative activity and the performance of UK new technology based firms

We present an innovation value chain analysis for a representative sample of new technology based firms (NTBFs) in the UK. This involves determining which factors lead to the usage of different knowledge sources and the relationships that exist between those sources of knowledge; the effect that each knowledge source has on innovative activity; and how innovation outputs affect the performance of NTBFs. We find that internal (i.e. R&D) and external knowledge sources are complementary for NTBFs, and that supply chain linkages have both a direct and indirect effect on innovation. NTBFs' skill resources matter throughout the innovation value chain, being positively associated with external knowledge linkages and innovation success, and also having a direct effect on growth independent of the effect on innovation. ©2010 IEEE.

Knowledge sources, innovative activity and the performance of UK new technology based firms

We present an innovation value chain analysis for a representative sample of new technology based firms (NTBFs) in the UK. This involves determining which factors lead to the usage of different knowledge sources and the relationships that exist between those sources of knowledge; the effect that each knowledge source has on innovative activity; and how innovation outputs affect the performance of NTBFs. We find that internal (i.e. R&D) and external knowledge sources are complementary for NTBFs, and that supply chain linkages have both a direct and indirect effect on innovation. NTBFs' skill resources matter throughout the innovation value chain, being positively associated with external knowledge linkages and innovation success, and also having a direct effect on growth independent of the effect on innovation. ©2010 IEEE.

Prospects and Opportunities. New Technology Trends yield new opportunities for Libraries and Information Centers

A number of important trends currently impact libraries. Academic libraries face the fundamental shift of collections toward ever increasing proportions of electronic content; public libraries continue to see vigorous interest in print materials, now supplemented by demand to provide e-books for lending. Breeding will explore these and other trends and describe some of the technologies available and emerging to help libraries meet the challenges involved in this context.

Factors affecting the adoption of new technology: The case of 311 government call centers

Government call centers (311) were first created to reduce the volume of non-emergency calls that were being placed to emergency 911 call centers. The number of 311 call centers increased from 57 in 2008 to about 300 in 2013. Considering that there are over 2,700 municipal government units across the United States, the adoption rate of the 311 centers is arguably low in the country. This dissertation is an examination of the adoption of 311 call centers by municipal governments. My focus is specifically on why municipal governments adopt 311 and identifying which barriers result in the non-adoption of 311 call centers. This dissertation is possibly the first study to examine the adoption of 311 call centers in the United States. ^ The dissertation study has identified several significant factors in the adoption and non-adoption of 311 government call centers. The following factors were significant in the adoption of 311 government call centers: managerial support, financial constraints, organizational responsiveness, strategic plan placement, and technology champion. The following factors were significant barriers that resulted in the non-adoption of a 311 government call center; no demand from citizens, start up costs, annual operating costs, unavailability of funding, and no obvious need for one. ^ If local government entities that do not have a 311 government call center decide to adopt one, this study will help them identify the conditions that need to be in place for successful adoption to occur. Local government officials would first need to address the barriers in setting up the 311 call centers. ^

2.1 Lloyd Gaines close up image

Charles Hamilton Houston and other NAACP attorneys assembled in early October 1939 to take depositions in preparation for the hearing scheduled a week later in Columbia to determine whether the university had complied with the Gaines decision. Attorneys took depositions from all of the instructors of the new LU law school as their preparation for the court proceedings wound down. The deposition of Lloyd Gaines was next. Attorneys planned to ask Gaines whether he considered Lincoln to be as good of a law school as Missouri and whether he planned to enroll. Called for questioning, Gaines did not respond. He could not be located anywhere. Lloyd’s mother, Callie Gaines, recalled that in January her son “left here to go to Kansas City to make a speech. That’s the last I saw of him.” While in Kansas City, Gaines spoke at the Centennial Methodist Church. He also looked for work, but not finding any caught a train for Chicago, telling people in Kansas City that he would stay a few days and return home.

A handbook of information on New York state education law, regulations and decisions of the commissioner of education, for the guidance of school administrators and boards of education in central and union free school districts of New York state,

"Index to helpful New York State Education Department bulletins and pamphlets": p. [3] of cover.

Law a century of progress, 1835-1935. Contributions in celebration of the 100th anniversary of the founding of the School of law of New York University.

Includes bibliographical references and index.