The expression of the gene for Azurin from Alcaligenes denitrificans in E. coli : a thesis presented in partial fulfilment of the requirements for the degree of Master of Science in Biochemistry at Massey University

Azurin is a protein which funtions in electron transport and has been found to bind copper when it is expressed in its native bacterial host. In this thesis the azurin from Alcaligenes denitrificans was used. This protein is 129 amino acids long with a molecular weight of 14. 600 dalions. The azurin coding gene from Alcaligenes denitrificans had previously been cloned into a plasmid which allows an E. coli expression system to be used. Azurin was purified from the E. coli hosts using the same procedures as for purifying copper-azurin from the native hosts but was found to remain apparently impure, according to spectrophotomctric data. Efforts to increase the production of the protein by using different expression systems and by refining the existing expression system failed to increase the apparent yield of copper-azurin. Efforts to refine the purification procedure also failed to increase the amount of copper-azurin that was purified. Various experiments were performed to demonstrate that azurin was expressed and processed correctly in the E. coli host. Protein was expressed in a copper-rich and copper-sparse environment. Copper-azurin was purified from the copper-rich environment, while very little copper-azurin could be extracted from the copper-sparse environment. The results described in this thesis suggest that when azurin from A. denitrificans is expressed in an E. coli host using standard media with no copper added, the predominant form of azurin produced in zinc-azunn. As mutants are going to made of this protein, conditions where the protein would bind only copper were required. The ideal conditions for this are still to be calculated but results from this thesis would suggest that copper concentrations in the region of 0.25 mM lead to 65% incorporation of copper. compared to 17% when no copper is added to the E. coli growth medium. E. coli cells were shown to grow with no apparent inhibition of growth in 3.0 mM of CuSO4. This concentration of copper in the growth medium may allow the production of a much higher ratio of copper-azurin compared to zinc-azurin than has been achieved so far

An overview of a problem in Hospitaller naval historiography

The Sovereign Military Hospitaller Order of St. John of Jerusalem of Rhodes and of Malta, frequently referred to as the Religion or the Hospital, is a particular institution. Recognised by Pope Paschal II in 1113, this centuries old Order had a double raison d'etre: To offer hospitality to Christian pilgrims to Jerusalem and to pursue the Holy War against the Infidel. However, the fall of Acre to the Saracens in 1291, made it impossible for the Hospitallers to fulfill their original vocation. After the capture and transformation of Rhodes into their base, they had to find a way of justifying their existence. They quickly came up with a characteristic answer - to wage naval war against Islam. So successful did the Knights become in this new role that the marine branch of the Order's military activities was to come to the greatest prominence, in Rhodes and later in Malta. What were the characteristics of the Order's navy? Small numbers, excellent design and an enviable tradition became the chief qualities of this small but effective nary. What, however, did the Hospitallers do with their galleys? Did they wage war on I slam? Or did their activities degenerate into privateering? This issue is hotly debated by historians of the Order. European Historians have a habit of shuddering away from accusing the Hospitallers of indulging in the corso, as corsairing is known in this context. This paper examines the topic in an attempt to unravel what has become a veritable Gordian knot.peer-reviewe

Borrower Self-Selection, Underwriting Costs, and Subprime Mortgage Credit Supply

In the U.S., households participate in two very different types of credit markets. Personal lending is characterized by continuous risk-based pricing in which lenders offer households a continuous distribution of borrowing possibilities based on estimates of their creditworthiness. This contrasts sharply with mortgage markets where lenders specialize in specific risk categories of borrowers and mortgage supply is stepwise linear. The contrast between continuous lending for personal loans and discrete lending by specialized lenders for mortgage credit has led to concerns regarding the efficiency and equity of mortgage lending. This paper sheds both theoretical and empirical light on the differences in the two credit markets. The theory section demonstrates why, in a perfectly competitive credit market where all lenders have the same underwriting technology, mortgage credit supply curves are stepwise linear and lenders specialize in prime or subprime lending. The empirical section then provides evidence that borrowers are being effectively sorted based on risk characteristics by the market

Exploring the potential of the union equality representative

Purpose – The purpose of this paper is to report on a qualitative study of the views of a cross-section of union regional activists and officers on the potential of a newly developed equality representative role in promoting greater equality in the workplace. Design/methodology/approach – Findings are based on an analysis of interview data incorporating the views and experiences of 32 officers and activists currently promoting equality in a region of the UK. Findings – Findings suggest that this is an equality initiative whose time may have come. This conclusion, however, is tempered by respondents' belief that the equality representative project can only fulfil its potential if the role is mainstreamed into branch structures, policies and processes and, crucially, that the same statutory recognition enjoyed by other union representatives is granted to the equality representative. Research limitations/implications – The project is at an early stage of development. Therefore, findings are based on the views of significant proponents of union equality initiatives in the region of study. Further research is needed to assess the actual impact of the new representatives as they become active in their branches and organisations. Practical implications – There are clear implications for management in terms of allowing representatives facility time and thus sharing the potential benefits of this new source of equality expertise. For the unions, the challenge is to mainstream the new role at all levels of the union and negotiate adequate facility time for the equality representative. Originality/value – The research is one of the few studies focussing on this new union strategy for better addressing the equality needs of their members and offers a clear insight into its potential success