Information System for the Transport Company

Cílem této práce je návrh systému a vytvoření funkční aplikace, která bude obsahovat nástroje sloužící pro evidenci a logistické řízení přepravní společnosti. V úvodní části je provedena rešerše systémů, které se zabývají touto problematikou a porovnání s nově navrhovaným systémem, který je předmětem této práce. Dále jsou popsány technologie a principy používané pro vývoj webových aplikací. Praktická část pojednává o vytvořeném informačním systému, funkcích a algoritmech použitých pro implementaci. Aplikace využívá skriptovací jazyk PHP a databázi MySQL.The main object of this diploma thesis is to design a system and an application utility for registering and logistic management of a transfer company. An opening part of this thesis includes an exploration of the facts about systems considering this problems and comparing these systems with the recently proposed system as the object of this thesis. Next part describes a technology and principles used for the web application design. The practical part of the thesis discusses recently designed information system, especially its functions and algorithms used for the implementation. An application is based on PHP script and MySQL database system.Fakulta elektrotechniky a informatik

Information system for service station

Cílem této práce je vytvoření funkční aplikace, která bude obsahovat nástroje sloužící pro zajištění činnosti autoservisu. V úvodní části je nutno provést rešerši systémů, které se zabývají touto problematikou. Rešerši je nutné doplnit o porovnání s nově navrhovaným systémem, který bude předmětem této práce. Úvodní část musí obsahovat analýzu navrhovaného řešení, která bude obsahovat popis použitých technologií, návrh databáze a aplikačního řešení. Pro vytvoření aplikace bude využit skriptovací jazyk PHP nebo JAVA a databáze Oracle nebo MySQL.An object of this thesis is to generate an application utility of tools serving the purpose of a car service station. An opening part of this thesis is to implement an exploration of arrangement engaged in these problems. A background research must be compared with a recently designed system as an object of this work. There must be a design analysis including the applied technics description as well as a database and an application project in the opening part. PHP and JAVA script and Oracle and MySQL databases will be used to create the aplication.Katedra informačních technologiíCílem práce bylo vytvoření funkční aplikace, která bude obsahovat nástroje sloužící pro zajištění činnosti autoservisu. Bakalářská práce byla zpracována na požadované úrovni a její cíl byl splněn. Zkoušený zodpověděl všechny dotazy komise

The 9aaTAD Transactivation Domains: From Gal4 to p53

<div><p>The family of the Nine amino acid Transactivation Domain, 9aaTAD family, comprises currently over 40 members. The 9aaTAD domains are universally recognized by the transcriptional machinery from yeast to man. We had identified the 9aaTAD domains in the p53, Msn2, Pdr1 and B42 activators by our prediction algorithm. In this study, their competence to activate transcription as small peptides was proven. Not surprisingly, we elicited immense 9aaTAD divergence in hundreds of identified orthologs and numerous examples of the 9aaTAD species' convergence. We found unforeseen similarity of the mammalian p53 with yeast Gal4 9aaTAD domains. Furthermore, we identified artificial 9aaTAD domains generated accidentally by others. From an evolutionary perspective, the observed easiness to generate 9aaTAD transactivation domains indicates the natural advantage for spontaneous generation of transcription factors from DNA binding precursors.</p></div

The 9aaTAD Is Exclusive Activation Domain in Gal4

<div><p>The Gal4 protein is a well-known prototypic acidic activator that has multiple activation domains. We have previously identified a new activation domain called the nine amino acid transactivation domain (9aaTAD) in Gal4 protein. The family of the 9aaTAD activators currently comprises over 40 members including p53, MLL, E2A and other members of the Gal4 family; Oaf1, Pip2, Pdr1 and Pdr3. In this study, we revised function of all reported Gal4 activation domains. Surprisingly, we found that beside of the activation domain 9aaTAD none of the previously reported activation domains had considerable transactivation potential and were not involved in the activation of transcription. Our results demonstrated that the 9aaTAD domain is the only decisive activation domain in the Gal4 protein. We found that the artificial peptides included in the original Gal4 constructs were results of an unintended consequence of cloning that were responsible for the artificial transcriptional activity. Importantly, the activation domain 9aaTAD, which is the exclusive activation domain in Gal4, is also the central part of a conserved sequence recognized by the inhibitory protein Gal80. We propose a revision of the Gal4 regulation, in which the activation domain 9aaTAD is directly linked to both activation function and Gal80 mediated inhibition.</p></div

Functional and non-functional activation domains in Gal4 activator.

<p>The Gal4—LexA hybrid constructs (BTM116 backbone) were assayed in L40 strain for activation of transcription. To distinguish activity of the N- and C-terminal activation domains in the Gal4 protein, a set of the Gal4 constructs were generated. The average value of the β-galactosidase activities from three independent experiments is presented as a percentage of the reference with standard deviation (means and plusmn; SD; n = 3). We standardized all results to previously reported Gal4 construct HaY including merely the activation domain 9aaTAD with the activity set to 100% [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0169261#pone.0169261.ref034" target="_blank">34</a>]. The LexA is <i>E</i>.<i>coli</i> DNA binding domain and the Gal4 DBD is <i>S</i>.<i>cerevisiae</i> Gal4 DNA binding domain, both generally used for the generation of fusion hybrids. The regions of Gal4 protein in the constructs are noted and graphically presented. Single dot means end of protein sequence, tree points mean continuing of the sequence, which is no more shown.</p

C-terminus of the Gal4DBD domain (92–100 aa) works as a half side of the 9aaTAD domain.

<p>A serious concern was found for not real activation function of the Gal4 acidic domain. The artificial 9aaTADs in pRJR200 and pRJR213 constructs were generated accidentally by others and represent so called Gal4 acidic domain. Here we restricted essential part of the Gal4 acidic domain to the recognized functional 9aaTAD region. In this constructs, the functionally unrelated peptides from Gal4 region (840–857 aa) subsidised unintentionally for the second half site of 9aaTAD domain. Artificial 9aaTADs was generated by fusion of the Gal4DBD domain and a half of the 9aaTAD of Gal4. The part of the Gal4DBD domain (92–100 aa) represent first half site of the 9aaTAD domain and was use in constructs to demonstrate capability to generate artificial 9aaTADs by fusion with the second half site of the Gal4 9aaTAD domain.</p

p53 9aaTADs activate transcription as small peptides.

<p>The predicted 9aaTADs in p53 from different species were tested for activation of transcription in LexA hybrid constructs. Similarity of p53 with Gal4 and Sox18 are highlighted. The construct 9p53, labelled with asterisk, has lower expression level compared with other constructs (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0162842#pone.0162842.s001" target="_blank">S1 Fig</a>). Animal picture from Flickr: Lowland Streaked Tenrec, Mantadia, Madagascar, Author: Frank Vassen; Elephant, Author: Jon Mountjoy; Igel (Hedgehog), Author: Mi chaela. All pictures have Creative Commons Attribution 2.0 Generic license.</p

Predicted 9aaTADs in Msn2 family.

<p>Prediction result for 9aaTADs in Msn2 and Msn4 activators retrieved by our 9aaTAD prediction algorithms. The conservation and variability of the 9aaTADs in the Msn2 and Gal4 families are shown.</p

Schema of the Gal4 regulation.

<p>Competition of Gal4 inhibitor Gal80 with mediators of transcription for the 9aaTAD domain. The peptides from <i>S</i>.<i>c</i>.Gal4 and <i>K</i>.<i>l</i>.Gal9 (Gal4 ortholog) interacting with Gal80 are shown (structural data for peptides interaction at PDB accession code 3E1K and 3BTS). The positions of activation domains 9aaTAD in the Gal80 binding peptides are highlighted (14 amino acid long Gal4 region is needed for maximal activation of transcription, construct H577, including the nine amino acid long 9aaTAD motif and four adjacent amino acids to the N-terminus and one to the C-terminus of the activation domain 9aaTAD). The artificial activation domains AD-I, AD-II and AD-III are not more shown in this figure.</p

The activation domain AD-III did not activate transcription.

<p>The LexA-Gal4 hybrid constructs with activation domains AD-III, 9aaTAD and their artificial variants were assayed in L40 strain for activation of transcription. Gal4 DBD is Gal4 DNA binding domain. The red marks part of the artificial activation domain 9aaTAD (¹/₂ 9aaTAD) and the blue marks part of the natural activation domain 9aaTAD (¹/₂ 9aaTAD). Both these parts generated strong activators in the fusion with the Gal4 DNA binding domain, Gal4 DBD (83–100 aa), which served as other part of the artificial activation domain 9aaTAD (the second ¹/₂ 9aaTAD). The artefact explained previous cloning errors of the reported construct pRJR200 with activation domain AD-III [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0169261#pone.0169261.ref031" target="_blank">31</a>]. The impact of proximal amino acids on the function of the activation domain 9aaTAD was tested. The related constructs with and without activation domains AD-III and 9aaTAD are shown again to overview their activities (G81, G49, G59, G42, G405). The average value of the β-galactosidase activities from three independent experiments is presented as a percentage of the reference with standard deviation (means and plusmn; SD; n = 3). We standardized all results to previously reported Gal4 construct HaY including merely the activation domain 9aaTAD with the activity set to 100% [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0169261#pone.0169261.ref034" target="_blank">34</a>]. The regions of Gal4 protein in the constructs are noted and graphically presented. The artificial activation domain AD-II is not more shown in this figure. The protein sequences are partially given by amino acid single letter code. Single dot means end of protein sequence, tree points mean continuing of the sequence, which is not more shown.</p