ANALIZA UČINAKA ULASKA REPUBLIKE HRVATSKE U EUROPSKU MONETARNU UNIJU ZA VELIKA IZVOZNA PODUZEĆA : Diplomski rad

Približavanjem Republike Hrvatske Europskoj monetarnoj uniji postaje sve jasnije kako će u bližoj budućnosti hrvatska kuna biti zamjenjena za euro. Za uvođenje eura država najprije mora zadovoljiti Maastrichtske kriterije ili kriteriji konvergencije. Svaki sustav ima neke prednosti i nedostatke pa tako i pristupanje Europskoj monetarnoj uniji. Na primjeru Slovenije i Slovačke se moglo vidjeti kako se nakon pristupanja Europskoj monetarnoj uniji prednosti i nedostaci različito manifestiraju, ovisno o situaciji u zemlji. Postavlja se pitanje kako će navedene prednosti i nedostaci djelovati na hrvatsko poduzeće i hoće li u konačnici koristi od pristupanja biti veće od troškova. Poduzeće će od ulaska u Europsku monetarnu uniju imati prednosti u vidu manjih transakcijskih troškova prilikom trgovine s inozemstvom, eliminacije valutnog rizika koje može potencijalno izazvati velike troškove fluktuacijom tečaja na tržištu jer je velika količina imovine i obveze vezana uz euro, te putem povećanja transparentnosti cijena imati će lakši zadatak pri odabiru povoljnih dobavljača. Od negativnih strana pristupanja Europskoj monetarnoj uniji za poduzeće mogu se izdvojiti troškovi prilagodbe informatičkih sustava i opreme, te obuka osoblja za korištenje te opreme, čiji troškovi su jednokratni i mogu se minimalizirati pravovremenom prilagodbom. Pravi učinak pristupanja moći će se ispravno analizirati nekoliko godina nakon pristupanja, no na temelju analize i ispitivanja provedenih na dostupnim podacima poduzeća može se zaključiti kako će poduzeće imati više koristi od troškova u poslovanju pristupanjem Republike Hrvatske Europskoj monetarnoj uniji.It is becoming clearer that the Republic of Croatia approaching the European monetary union in the near future euro will supstitute the Croatian kuna. Before the implementation of euro a contry must first fulfill the Maastricht criteria also known as criteria of convergence. All systems have some advantages and disadvantages, and so does joining the European monetary union. The examples of Slovenia and Slovakia show that those advantages and disadvantages present themself diferently after joining the union, depending on the conditions in the contry. The question is how will the advantages and disatvantages effect Croatian companys and will joining the European monetary union bring more benefit or cost. The company will after accession to the European monetary union expirience advantages in lower transaction costs in international trade, elimination of currency risk that can cause large costs by currency fluctuations when assets and obligations are tied to euro. By larger transperency in prices the company will have an easier task in selecting suppliers. Of the disadvantages of joining the European monetary union, the compay can expect to expirience an raise in costs for adjusting its computer systems and gear, as well as training its staff for using the systems and gears. These disatvantagese are one-time costs and can be minimalised by adequate preparation. The real effects of joining can be properly analized several years after European monetary union membership, but on the basis of analizing and examination on available information of the company, it is concluded that the company will have more benefit then cost from joining the European monetary union

ANALIZA UČINAKA ULASKA REPUBLIKE HRVATSKE U EUROPSKU MONETARNU UNIJU ZA VELIKA IZVOZNA PODUZEĆA : Diplomski rad

Približavanjem Republike Hrvatske Europskoj monetarnoj uniji postaje sve jasnije kako će u bližoj budućnosti hrvatska kuna biti zamjenjena za euro. Za uvođenje eura država najprije mora zadovoljiti Maastrichtske kriterije ili kriteriji konvergencije. Svaki sustav ima neke prednosti i nedostatke pa tako i pristupanje Europskoj monetarnoj uniji. Na primjeru Slovenije i Slovačke se moglo vidjeti kako se nakon pristupanja Europskoj monetarnoj uniji prednosti i nedostaci različito manifestiraju, ovisno o situaciji u zemlji. Postavlja se pitanje kako će navedene prednosti i nedostaci djelovati na hrvatsko poduzeće i hoće li u konačnici koristi od pristupanja biti veće od troškova. Poduzeće će od ulaska u Europsku monetarnu uniju imati prednosti u vidu manjih transakcijskih troškova prilikom trgovine s inozemstvom, eliminacije valutnog rizika koje može potencijalno izazvati velike troškove fluktuacijom tečaja na tržištu jer je velika količina imovine i obveze vezana uz euro, te putem povećanja transparentnosti cijena imati će lakši zadatak pri odabiru povoljnih dobavljača. Od negativnih strana pristupanja Europskoj monetarnoj uniji za poduzeće mogu se izdvojiti troškovi prilagodbe informatičkih sustava i opreme, te obuka osoblja za korištenje te opreme, čiji troškovi su jednokratni i mogu se minimalizirati pravovremenom prilagodbom. Pravi učinak pristupanja moći će se ispravno analizirati nekoliko godina nakon pristupanja, no na temelju analize i ispitivanja provedenih na dostupnim podacima poduzeća može se zaključiti kako će poduzeće imati više koristi od troškova u poslovanju pristupanjem Republike Hrvatske Europskoj monetarnoj uniji.It is becoming clearer that the Republic of Croatia approaching the European monetary union in the near future euro will supstitute the Croatian kuna. Before the implementation of euro a contry must first fulfill the Maastricht criteria also known as criteria of convergence. All systems have some advantages and disadvantages, and so does joining the European monetary union. The examples of Slovenia and Slovakia show that those advantages and disadvantages present themself diferently after joining the union, depending on the conditions in the contry. The question is how will the advantages and disatvantages effect Croatian companys and will joining the European monetary union bring more benefit or cost. The company will after accession to the European monetary union expirience advantages in lower transaction costs in international trade, elimination of currency risk that can cause large costs by currency fluctuations when assets and obligations are tied to euro. By larger transperency in prices the company will have an easier task in selecting suppliers. Of the disadvantages of joining the European monetary union, the compay can expect to expirience an raise in costs for adjusting its computer systems and gear, as well as training its staff for using the systems and gears. These disatvantagese are one-time costs and can be minimalised by adequate preparation. The real effects of joining can be properly analized several years after European monetary union membership, but on the basis of analizing and examination on available information of the company, it is concluded that the company will have more benefit then cost from joining the European monetary union

CRISPR-Cas in Escherichia coli: regulation by H-NS, LeuO and temperature

CRISPR-Cas adaptive immune systems are present in many bacteria and archaea and provide protection against invading DNA such as phages and plasmids. These systems are very versatile and complex in their gene composition and genomic architecture. CRISPR-Cas systems are classified into 2 classes, 6 types and 33 subtypes although this number is not definitive and the research is ongoing. All CRISPR-Cas systems have been thoroughly investigated in order to better understand the mechanism of CRISPR immunity enabling its use as a tool in genome editing and other biotechnological applications. However, regulation of the CRISPR-Cas system is also very complex and still not fully understood; it must provide optimal protection without introducing harmful consequences to the host. In this review we give an overview on the regulation of the CRISPR-Cas system Class 1 Type I-E in Escherichia coli with the emphasis on the role of temperature in regulation of the CRISPR-Cas activity and the interplay of the key regulators H-NS and StpA repressors and LeuO antirepressor in regulation of cas gene expression and HtpG chaperone in maintaining functional levels of Cas3.</p

CRISPR-Cas adaptation in Escherichia coli requires RecBCD helicase but not nuclease activity, is independent of homologous recombination, and is antagonized by 5' ssDNA exonucleases

© The Author(s) 2018. Prokaryotic adaptive immunity is established against mobile genetic elements (MGEs) by 'naïve adaptation' when DNA fragments from a newly encountered MGE are integrated into CRISPR-Cas systems. In Escherichia coli, DNA integration catalyzed by Cas1- Cas2 integrase is well understood in mechanistic and structural detail butmuch less is known about events prior to integration that generate DNA for capture by Cas1-Cas2. Naïve adaptation in E. coli is thought to depend on the DNA helicase-nuclease RecBCD for generating DNA fragments for capture by Cas1- Cas2. The genetics presented here show that naïve adaptation does not require RecBCD nuclease activity but that helicase activity may be important. RecA loading by RecBCD inhibits adaptation explaining previously observed adaptation phenotypes that implicated RecBCD nuclease activity. Genetic analysis of other E. coli nucleases and naïve adaptation revealed that 5' ssDNA tailed DNA molecules promote new spacer acquisition. We show that purified E. coli Cas1-Cas2 complex binds to and nicks 5' ssDNA tailed duplexes and propose that E. coli Cas1-Cas2 nuclease activity on such DNA structures supports naïve adaptation

The role of RecBCD enzyme and other nucleases in Escherichia coli CRISPR-Cas naïve adaptation

Sustav CRISPR-Cas je obrambeni mehanizam bakterija od stranih genetičkih elemenata. Za pružanje zaštite je neophodna ugradnja fragmenta strane molekule DNA (razmaknice) u lokus CRISPR. Ugradnja nove razmaknice se naziva naivnom adaptacijom, a za ugradnju je neophodan proteinski kompleks Cas1-Cas2. Nije poznato kako nastaju fragmenti za ugradnju, no smatra se da bitnu ulogu ima nukleazna aktivnost enzima RecBCD. U ovom radu je detaljnije istražena uloga enzima RecBCD i drugih egzonukleaza u naivnoj adaptaciji. Praćenjem učinkovitosti adaptacije in vivo u sojevima E. coli mutiranim za istraživane gene je pokazano da nukleazna aktivnost enzima RecBCD nije neophodna, već samo helikazna, te da nastajanje 5'-stršećih krajeva DNA pomoću egzonukleaza pomaže adaptaciju. Utvrđeno je in vitro da kompleks Cas1-Cas2 može cijepati DNA s 5'-stršećim krajevima, a sekvenciranjem je utvrđeno da većina ugrađenih razmaknica potječe s kromosoma. Rezultati potvrđuju ulogu enzima RecBCD u stvaranju fragmenata DNA, a sam proces adaptacije ne utječe na vijabilnost.CRISPR-Cas system is a bacterial defence mechanism against foreign genetic elements based on insertion of foreign DNA fragments (spacers) into the CRISPR array. Acquisition of new spacers is called naïve adaptation and is catalysed by Cas1-Cas2 protein complex. It is unknown how fragments for insertion are generated, but it is believed that the nuclease activity of RecBCD enzyme plays an important role. The aim of this research was to further elucidate the role of RecBCD and other exonucleases in naïve adaptation in E. coli. In vivo adaptation efficiency measurements showed that the helicase activity of RecBCD is essential and that generation of 5' overhangs by host exonucleases promotes adaptation. Cas1-Cas2 complex cleaved DNA with 5' overhangs in vitro and sequencing analysis showed that the majority of new spacers were of chromosomal origin. Overall, results confirm RecBCD's involvement in naïve adaptation with no detrimental effects on host viability

The role of RecBCD enzyme and other nucleases in Escherichia coli CRISPR-Cas naïve adaptation

Sustav CRISPR-Cas je obrambeni mehanizam bakterija od stranih genetičkih elemenata. Za pružanje zaštite je neophodna ugradnja fragmenta strane molekule DNA (razmaknice) u lokus CRISPR. Ugradnja nove razmaknice se naziva naivnom adaptacijom, a za ugradnju je neophodan proteinski kompleks Cas1-Cas2. Nije poznato kako nastaju fragmenti za ugradnju, no smatra se da bitnu ulogu ima nukleazna aktivnost enzima RecBCD. U ovom radu je detaljnije istražena uloga enzima RecBCD i drugih egzonukleaza u naivnoj adaptaciji. Praćenjem učinkovitosti adaptacije in vivo u sojevima E. coli mutiranim za istraživane gene je pokazano da nukleazna aktivnost enzima RecBCD nije neophodna, već samo helikazna, te da nastajanje 5'-stršećih krajeva DNA pomoću egzonukleaza pomaže adaptaciju. Utvrđeno je in vitro da kompleks Cas1-Cas2 može cijepati DNA s 5'-stršećim krajevima, a sekvenciranjem je utvrđeno da većina ugrađenih razmaknica potječe s kromosoma. Rezultati potvrđuju ulogu enzima RecBCD u stvaranju fragmenata DNA, a sam proces adaptacije ne utječe na vijabilnost.CRISPR-Cas system is a bacterial defence mechanism against foreign genetic elements based on insertion of foreign DNA fragments (spacers) into the CRISPR array. Acquisition of new spacers is called naïve adaptation and is catalysed by Cas1-Cas2 protein complex. It is unknown how fragments for insertion are generated, but it is believed that the nuclease activity of RecBCD enzyme plays an important role. The aim of this research was to further elucidate the role of RecBCD and other exonucleases in naïve adaptation in E. coli. In vivo adaptation efficiency measurements showed that the helicase activity of RecBCD is essential and that generation of 5' overhangs by host exonucleases promotes adaptation. Cas1-Cas2 complex cleaved DNA with 5' overhangs in vitro and sequencing analysis showed that the majority of new spacers were of chromosomal origin. Overall, results confirm RecBCD's involvement in naïve adaptation with no detrimental effects on host viability

ANALIZA UČINAKA ULASKA REPUBLIKE HRVATSKE U EUROPSKU MONETARNU UNIJU ZA VELIKA IZVOZNA PODUZEĆA : Diplomski rad

Približavanjem Republike Hrvatske Europskoj monetarnoj uniji postaje sve jasnije kako će u bližoj budućnosti hrvatska kuna biti zamjenjena za euro. Za uvođenje eura država najprije mora zadovoljiti Maastrichtske kriterije ili kriteriji konvergencije. Svaki sustav ima neke prednosti i nedostatke pa tako i pristupanje Europskoj monetarnoj uniji. Na primjeru Slovenije i Slovačke se moglo vidjeti kako se nakon pristupanja Europskoj monetarnoj uniji prednosti i nedostaci različito manifestiraju, ovisno o situaciji u zemlji. Postavlja se pitanje kako će navedene prednosti i nedostaci djelovati na hrvatsko poduzeće i hoće li u konačnici koristi od pristupanja biti veće od troškova. Poduzeće će od ulaska u Europsku monetarnu uniju imati prednosti u vidu manjih transakcijskih troškova prilikom trgovine s inozemstvom, eliminacije valutnog rizika koje može potencijalno izazvati velike troškove fluktuacijom tečaja na tržištu jer je velika količina imovine i obveze vezana uz euro, te putem povećanja transparentnosti cijena imati će lakši zadatak pri odabiru povoljnih dobavljača. Od negativnih strana pristupanja Europskoj monetarnoj uniji za poduzeće mogu se izdvojiti troškovi prilagodbe informatičkih sustava i opreme, te obuka osoblja za korištenje te opreme, čiji troškovi su jednokratni i mogu se minimalizirati pravovremenom prilagodbom. Pravi učinak pristupanja moći će se ispravno analizirati nekoliko godina nakon pristupanja, no na temelju analize i ispitivanja provedenih na dostupnim podacima poduzeća može se zaključiti kako će poduzeće imati više koristi od troškova u poslovanju pristupanjem Republike Hrvatske Europskoj monetarnoj uniji.It is becoming clearer that the Republic of Croatia approaching the European monetary union in the near future euro will supstitute the Croatian kuna. Before the implementation of euro a contry must first fulfill the Maastricht criteria also known as criteria of convergence. All systems have some advantages and disadvantages, and so does joining the European monetary union. The examples of Slovenia and Slovakia show that those advantages and disadvantages present themself diferently after joining the union, depending on the conditions in the contry. The question is how will the advantages and disatvantages effect Croatian companys and will joining the European monetary union bring more benefit or cost. The company will after accession to the European monetary union expirience advantages in lower transaction costs in international trade, elimination of currency risk that can cause large costs by currency fluctuations when assets and obligations are tied to euro. By larger transperency in prices the company will have an easier task in selecting suppliers. Of the disadvantages of joining the European monetary union, the compay can expect to expirience an raise in costs for adjusting its computer systems and gear, as well as training its staff for using the systems and gears. These disatvantagese are one-time costs and can be minimalised by adequate preparation. The real effects of joining can be properly analized several years after European monetary union membership, but on the basis of analizing and examination on available information of the company, it is concluded that the company will have more benefit then cost from joining the European monetary union