Recombining overlapping BACs into a single larger BAC

BACKGROUND: BAC clones containing entire mammalian genes including all the transcribed region and long range controlling elements are very useful for functional analysis. Sequenced BACs are available for most of the human and mouse genomes and in many cases these contain intact genes. However, large genes often span more than one BAC, and single BACs covering the entire region of interest are not available. Here we describe a system for linking two or more overlapping BACs into a single clone by homologous recombination. RESULTS: The method was used to link a 61-kb insert carrying the final 5 exons of the human CFTR gene onto a 160-kb BAC carrying the first 22 exons. Two rounds of homologous recombination were carried out in the EL350 strain of bacteria which can be induced for the Red genes. In the first round, the inserts of the two overlapping BACs were subcloned into modified BAC vectors using homologous recombination. In the second round, the BAC to be added was linearised with the very rare-cutting enzyme I-PpoI and electroporated into recombination efficient EL350 bacteria carrying the other BAC. Recombined BACs were identified by antibiotic selection and PCR screening and 10% of clones contained the correctly recombined 220-kb BAC. CONCLUSION: The system can be used to link the inserts from any overlapping BAC or PAC clones. The original orientation of the inserts is not important and desired regions of the inserts can be selected. The size limit for the fragments recombined may be larger than the 61 kb used here and multiple BACs in a contig could be combined by alternating use of the two pBACLink vectors. This system should be of use to many investigators wishing to carry out functional analysis on large mammalian genes which are not available in single BAC clones

The social identity of the Post - Olympic use of the Olympic Village settlement in Athens-Greece

The Olympic Village settlement was constructed under the responsibility of Workers Housing Organization during the preparation of the 2004 Olympic Games. It is a new city with 2.292 housing units which was developed in a very short time period (3.5 years) via the system of organized planning. It is situated in an area outside of the urban fabric of Attica and the post - Olympic settlement has a population of approximately 10,000 inhabitants with different demographic, socioeconomic and cultural characteristics. The inhabitants of the post - Olympic settlement are beneficiaries of the Workers Housing Organization. They have been selected after a lottery and satisfied previously determined social criteria. The methodology of the research was supported by the analysis and statistical process of 2,292 questionnaires supplemented to the beneficiaries by the year 2003, during the Census of Workers Housing Organization for the lottery of housing units of Olympic Village settlement. The results of the study determine the social identity of the post-Olympic Village after re-designing the data base and analyzing main characteristics of the inhabitants such as the synthesis of the households, the age, the labor status, the income and the educational level of the population. The analysis aims to contribute to the identification of the resident's needs which is an essential factor for the successful post-Olympic use of the settlement

Identification of Marine Biotechnology Value Chains with High Potential in the Northern Mediterranean Region

© 2023. The authors. This document is made available under the CC-BY 4.0 license http://creativecommons.org/licenses/by /4.0/ This document is the Accepted version of a Published Work that appeared in final form in Marine Drugs. To access the final edited and published work see https://doi.org/10.3390/md21070416Marine (blue) biotechnology is an emerging field enabling the valorization of new products and processes with massive potential for innovation and economic growth. In the Mediterranean region, this innovation potential is not exploited as well as in other European regions due to a lack of a clear identification of the different value chains and the high fragmentation of business innovation initiatives. As a result, several opportunities to create an innovative society are being missed. To address this problem, eight Northern Mediterranean countries (Croatia, France, Greece, Mar. Drugs 2023, 21, 416. https://doi.org/10.3390/md21070416 https://www.mdpi.com/journal/marinedrugs Mar. Drugs 2023, 21, 416 2 of 26 Italy, Montenegro, Portugal, Slovenia and Spain) established five national blue biotechnology hubs to identify and address the bottlenecks that prevent the development of marine biotechnology in the region. Following a three-step approach (1. Analysis: setting the scene; 2. Transfer: identification of promising value chains; 3. Capitalization: community creation), we identified the three value chains that are most promising for the Northern Mediterranean region: algae production for added-value compounds, integrated multi-trophic aquaculture (IMTA) and valorization aquaculture/fisheries/processing by-products, unavoidable/unwanted catches and discards. The potential for the development and the technical and non-technical skills that are necessary to advance in this exciting field were identified through several stakeholder events which provided valuable insight and feedback that should be addressed for marine biotechnology in the Northern Mediterranean region to reach its full potential

Identification of Marine Biotechnology Value Chains with High Potential in the Northern Mediterranean Region

©2023. This manuscript version is made available under the CC-BY 4.0 license http://creativecommons.org/licenses/by/4.0/ This document is the Published, version of a Published Work that appeared in final form in Marine Drugs. To access the final edited and published work see https://doi.org/ 10.3390/md21070416Marine (blue) biotechnology is an emerging field enabling the valorization of new products and processes with massive potential for innovation and economic growth. In the Mediterranean region, this innovation potential is not exploited as well as in other European regions due to a lack of a clear identification of the different value chains and the high fragmentation of business innovation initiatives. As a result, several opportunities to create an innovative society are being missed. To address this problem, eight Northern Mediterranean countries (Croatia, France, Greece Italy, Montenegro, Portugal, Slovenia and Spain) established five national blue biotechnology hubs to identify and address the bottlenecks that prevent the development of marine biotechnology in the region. Following a three-step approach (1. Analysis: setting the scene; 2. Transfer: identifi cation of promising value chains; 3. Capitalization: community creation), we identified the three value chains that are most promising for the Northern Mediterranean region: algae production for added-value compounds, integrated multi-trophic aquaculture (IMTA) and valorization aquacul ture/fisheries/processing by-products, unavoidable/unwanted catches and discards. The potential for the development and the technical and non-technical skills that are necessary to advance in this exciting field were identified through several stakeholder events which provided valuable insight and feedback that should be addressed for marine biotechnology in the Northern Mediterranean region to reach its full potential

Construction of mammalian artificial chromosome vectors for expression of therapeutic genes

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Growth, feed utilization, health and organoleptic characteristics of European seabass (Dicentrarchus labrax) fed extruded diets including low and high levels of three different legumes

Three legumes [field peas (P), chickpeas (CP) and faba beans (B)] were evaluated at two inclusion levels; 170 (L) and 350 g kg- 1 (H) in a 14-week experiment with triplicate groups of 97.9 ñ 6.1 g European seabass. A control diet included wheat meal, fish meal (FM) and a mixture of plant ingredients as protein sources. Diets, processed in a twin-screw extruder, were isonitrogenous and isoenergetic. All experimental diets improved growth compared to the control. Weight gain, feed intake, FCR and SGR were improved for fish fed diet CPL, while fish fed diet CPH gave higher FCR. Protein and starch digestibility were highest for the control diet, while fish fed diets CPL and PL showed significantly lower protein ADCs and lower starch ADC (P = 0.05) for diet PH. Among H diets, significantly better fillet yield was found for fish fed diet PH, but no other differences were found in seabass fillet organoleptic characteristics. Serum glucose and cholesterol were elevated in fish fed CP diets, while protein and triacylglycerols did not show significant differences among treatments. Fish fed the test diets showed no significant evidence of either immunosuppression or immunostimulation. Histology of liver, spleen, kidney and foregut revealed no pathological abnormalities. Field peas, chickpeas and faba beans can be included in European seabass diets up to 350 g kg- 1 substituting for wheat with no negative effects on growth performance, carcass composition or organoleptic characteristics