An in vitro transposon system for highly regulated gene expression: construction of Escherichia coli strains with arabinose-dependent growth at low temperatures.

Placing a gene of interest under the control of an inducible promoter greatly aids the purification, localization and functional analysis of proteins but usually requires the sub-cloning of the gene of interest into an appropriate expression vector. Here, we describe an alternative approach employing in vitro transposition of Tn Omega P(BAD) to place the highly regulable, arabinose inducible P(BAD) promoter upstream of the gene to be expressed. The method is rapid, simple and facilitates the optimization of expression by producing constructs with variable distances between the P(BAD) promoter and the gene. To illustrate the use of this approach, we describe the construction of a strain of Escherichia coli in which growth at low temperatures on solid media is dependent on threshold levels of arabinose. Other uses of the transposable promoter are also discussed.

The open parliament in the age of the internet : can the people now collaborate with legislatures in lawmaking?

This work seeks to address those questions and evaluate other international experiences and experiments designed to achieve the same ends. The book is based on a study of two particular cases where parliamentary bodies designed and implemented participatory digital processes, namely, the e-Democracy Program developed by the Brazilian House of Representatives, and the Virtual Senator Program developed by the Chilean Senate. The text unfolds in the form of a systematic analysis of institutional aspects embracing political and organizational elements as well as the social aspects associated to the application of digital democracy in parliaments. The investigation shows that at the stage they found themselves in 2010 those projects had only brought in very incipient results in regard to the aspects of enhancing representativity in decision making processes, aggregating collective intelligence to the legislative process or transparency to parliamentary performances, even though all of those are precious components of any democracy that deems itself to be participatory and deliberative. Nevertheless, such experiences have had the merit of contributing towards the gradual construction of more effective participatory mechanisms, complementary to the political representation system in place.

Collective evolution characteristics and computer simulation of short fatigue cracks

Fatigue testing was conducted using a kind of triangular isostress specimen to obtain the short-fatigue-crack behaviour of a weld low-carbon steel. The experimental results show that short cracks continuously initiate at slip bands within ferrite grain domains and the crack number per unit area gradually increases with increasing number of fatigue cycles. The dispersed short cracks possess an orientation preference, which is associated with the crystalline orientation of the relevant slip system. Based on the observed collective characteristics, computer modelling was carried out to simulate the evolution process of initiation, propagation and coalescence of short cracks. The simulation provides progressive displays which imitate the appearance of experimental observations. The results of simulation indicate that the crack path possesses a stable value of fractal dimension whereas the critical value of percolation covers a wide datum band, suggesting that the collective evolution process of short cracks is sensitive to the pattern of crack site distribution.

Proposal for improving fishing opportunities off Pt. Loma by construction of an artificial reef

(Document pdf contains 42 pages)

Pond construction: selecting good places for ponds

(4pp.)

Pond construction: design and layout of ponds

(4pp.)