Optimization of the Expression of 1,3-1,4-β-glucanase Gene from Rhizomucor miehei in the Komagataella kurtzmanii yeast

2019 ◽  
Vol 35 (5) ◽  
pp. 3-11 ◽  
Author(s):  
I.I. Gubaidullin ◽  
A.S. Fedorov ◽  
D.G. Kozlov
Keyword(s):  
Target Gene ◽  
Rhizomucor Miehei ◽  
Leader Peptide ◽  
Promoter Regions ◽  
Functional Elements ◽  
Resource Center ◽  
The Status ◽  
Pichia Pastoris Yeast ◽  
The Difference ◽  
Genetic Constructs

Key functional elements of the vector (promoter, leader and terminator regions) that provide the expression of a target l,3-l,4-(3-glucanase gene from Rhizomucor miehei in the Komagataella kurtzmanii yeast have been optimized. It was shown that the promoter regions of the gene AOX1 from the Pichia pastoris yeast currently reclassified as Komagataella phaffti and from К. kurtzmanii yeast as parts of a vector provided equal levels of expression of the target gene in the cells of the recipient strain К. kurtzmanii Y727his4, i.e. they were completely interchangeable. This means that genetic constructs that were previously developed for the biosynthesis of recombinant proteins in К. phajfii are able to provide an effective expression in the К kurtzmanii yeast. The leader peptide MF4I (used as a variant of mif4I containing one amino acid substitution) and the leader peptide maxHH (containing the double proregion of the Hspl50 protein from Saccharomyces cerevisiae) confirmed the status of the most powerful elements among the five leader sequences analyzed. Their efficiency was 1.7 times higher than that of the standard leader from the yeast alpha-factor, and by 20% higher than the characteristics of the second group of artificial leaders. At the same time, it was found that, the choice of the terminator region had the strongest influence on the expression of the target gene among all of the vector functional elements. The best terminator elements were variants derived from the transcription termination region of the AOX1 gene, and the difference in the expression level of the target gene using different terminators was approximately 4.5 times. Based on the analysis of the obtained data, the optimal composition of the key functional elements of the expression vector was determined ; it included the promoter and terminator regions of the AOX1 yeast gene and one of the artificial leaders, mif4I or maxHH. β-glucanase, Komagataella kurtzmanii, yeast, secretion, strain producer The work was financially supported by the Ministry of Science and Higher education of the Russian Federation (Unique Project Identifier RFMEFI60717X0179) using the Unique Scientific Facility of the National Bio-Resource Center «All-Russian Collection of Industrial Microorganisms», NRC «Kurchatov Institute» - GOSNIIGENETIKA

Increase in the Production of Endo-1,4-β-Xylanase from Paenibacillus brasilensis in Pichia pastoris

2019 ◽  
Vol 35 (6) ◽  
pp. 30-38 ◽  
Author(s):  
L.N. Borshchevskaya ◽  
T.L. Gordeeva ◽  
S.P. Sineoky
Keyword(s):  
Pichia Pastoris ◽  
Target Gene ◽  
Heterologous Protein ◽  
Ministry Of Education ◽  
Kurchatov Institute ◽  
Xylanase Production ◽  
Resource Center ◽  
Recombination System ◽  
Codon Composition ◽  
Pichia Pastoris Yeast

A Pichia pastoris yeast strain producing endo-l,4-β-xylanase from Paenibacillus brasilensis with an activity of 54,400 U/mL after 140 h of fermentation in a laboratory fermenter has been obtained. A number of approaches were used to increase the level of the xylanase production in this strain: optimization of the target gene codon composition, multiple integration of the expression cassette into the recipient strain chromosome using the Cre-lox recombination system, and also improving the heterologous protein folding via the overexpression of the HAC1i gene from Pichia pastoris. xylanase, xylan, Cre-lox system, HAC1p transcriptional activator, multicopy strain, Paenibacillus brasilensis, Pichia pastoris The work was performed with the financial support of the Ministry of Education and Science of Russia (Unique Project Identifier RFMEFI60717X0180) using the Multipurpose Scientific Installation of «All-Russian Collection of Industrial Microorganisms» National Bio-Resource Center, NRC «Kurchatov Institute» - GosNIIgenetika.

Motivasi dan kepuasan kerja pegawai puskesmas BLUD dan non-BLUD Kabupaten Semarang

2018 ◽  
Vol 2 (3) ◽  
pp. 111
Author(s):  
Aswindar Adhi Gumilang ◽  
Tri Pitara Mahanggoro ◽  
Qurrotul Aini
Keyword(s):  
Public Health ◽  
Job Satisfaction ◽  
Financial Management ◽  
Work Motivation ◽  
P Value ◽  
The Public ◽  
The Status ◽  
The Difference ◽  
Demand For Health ◽  
Public Demand

The public demand for health service professionalism and transparent financial management made some Puskesmas in Semarang regency changed the status of public health center to BLUD. The implementation of Puskesmas BLUD and non-BLUD requires resources that it can work well in order to meet the expectations of the community. The aim of this study is to know the difference of work motivation and job satisfaction of employees in Puskesmas BLUD and non-BLUD. Method of this research is a comparative descriptive with a quantitative approach. The object of this research are work motivation and job satisfaction of employees in Puskesmas BLUD and non-BLUD Semarang regency. This Research showed that Sig value. (P-value) work motivation variable was 0.019 smaller than α value (0.05). It showed that there was a difference of work motivation of employees in Puskemas BLUD and non-BLUD. Sig value (P-value) variable of job satisfaction was 0.020 smaller than α value (0.05). It showed that there was a difference of job satisfaction of BLUD and non-BLUD. The average of non-BLUD employees motivation were 76.59 smaller than the average of BLUD employees were 78.25. The average of job satisfaction of BLUD employees were 129.20 bigger than the average of non-BLUD employee were 124.26. Job satisfaction of employees in Puskesmas BLUD was higher than non-BLUD employees.

Vier Arten von Römern unter den Franken im 6. bis 8. Jh.

2016 ◽  
Vol 133 (1) ◽  
pp. 459-468
Author(s):  
Detlef Liebs
Keyword(s):  
Criminal Law ◽  
Civil Law ◽  
Roman Law ◽  
Procedural Law ◽  
The Status ◽  
The Difference ◽  
Law Texts ◽  
Fourth Kind

Abstract Four kinds of Romans in the Frankish kingdoms in the 6th to 8th centuries. Roman law texts from Merowingian Gaul make a difference between cives Romani, Latini and dediticii, all considered as Romans. This difference mattered only to slaves who had been freed. The status of Latin and dediticius was hereditary, whereas the descendants of one who had been freed as civis Romanus were free born Romans, who should be classified as a proper, a fourth kind of beeing Roman; it was the standard kind. The difference was important in civil law, procedural law and criminal law, especially in wergeld, the sum to be payed for expiation when somebody had been killed: Who had killed a Roman, had to pay different sums according to the status of the killed.

Through the Lens of Gadamer’s Hermeneutics: In comparison between Amina Wadud’s and Mohammed Talbi’s Interpretations of Q. 4:34

SUHUF ◽  
2017 ◽  
Vol 9 (2) ◽  
pp. 193-214
Author(s):  
Afifur Rochman Sya'rani
Keyword(s):  
Gender Inequality ◽  
Married Couple ◽  
Contextual Approach ◽  
The Status ◽  
Similarities And Differences ◽  
The Difference ◽  
Muslim Scholars ◽  
The Way

Most of traditional Muslim exegetes interpret Q. 4:34 in terms of maintaining the superiority of men over women. Some progressive Muslim scholars then insist a contextual approach to the verse to criticize gender inequality. Among some progressive Muslim scholars, this article comparatively examines the interpretations of Amina Wadud and Mohammed Talbi of Q. 4:34. Although both of them propose a contextual reading of the verse, they have different intellectual background, approach and method in interpreting the Qur’ān. The questions are to what extent the similarities and differences of both Wadud’s and Talbi’s interpretation of Q. 4:34 and how far their interpretations reflect their respective intention and perspective? Applying Gadamer’s hermeneutical approach, the article concludes that [1] Both Wadud and Talbi argue that the verse does not establish the superiority of men over women, but acknowledges duties division among married couple; [2] the difference among their interpretations is on the status of relationship among married couple; [3] Wadud’s and Talbi’s interpretations represent their respective hermeneutical situations and the way they define ontologically the nature of  interpretation and Qur’anic hermeneutics affect on producing the meanings of the verse.

scholarly journals What Difference Does Posthumanism Make?

2021 ◽  
pp. 1-7
Author(s):  
Rachel J. Crellin ◽  
Oliver J.T. Harris
Keyword(s):  
Productive Force ◽  
The World ◽  
The Status ◽  
The Difference ◽  
The Relationship

In this paper we argue that to understand the difference Posthumanism makes to the relationship between archaeology, agency and ontology, several misconceptions need to be corrected. First, we emphasize that Posthumanism is multiple, with different elements, meaning any critique needs to be carefully targeted. The approach we advocate is a specifically Deleuzian and explicitly feminist approach to Posthumanism. Second, we examine the status of agency within Posthumanism and suggest that we may be better off thinking about affect. Third, we explore how the approach we advocate treats difference in new ways, not as a question of lack, or as difference ‘from’, but rather as a productive force in the world. Finally, we explore how Posthumanism allows us to re-position the role of the human in archaeology,

Identification of an upstream regulatory sequence that mediates the transcription of mox genes in Methylobacterium extorquens AM1

Microbiology ◽  
2005 ◽  
Vol 151 (11) ◽  
pp. 3723-3728 ◽  
Author(s):  
Meng Zhang ◽  
Kelly A. FitzGerald ◽  
Mary E. Lidstrom
Keyword(s):  
Promoter Activity ◽  
Site Directed Mutagenesis ◽  
Regulatory Sequence ◽  
Methylobacterium Extorquens ◽  
Promoter Regions ◽  
Enhancer Element ◽  
Conserved Sequence ◽  
Normal Expression ◽  
Genetic Constructs ◽  
Transcriptional Fusions

A multiple A-tract sequence has been identified in the promoter regions for the mxaF, pqqA, mxaW, mxbD and mxcQ genes involved in methanol oxidation in Methylobacterium extorquens AM1, a facultative methylotroph. Site-directed mutagenesis was exploited to delete or change this conserved sequence. Promoter-xylE transcriptional fusions were used to assess promoter activity in these mutants. A fiftyfold drop in the XylE activity was observed for the mxaF and pqqA promoters without this sequence, and a five- to sixfold drop in the XylE activity was observed for the mxbD and mxcQ promoters without this sequence. Mutants were generated in the chromosomal copies in which this sequence was either deleted or altered, and these mutants were unable to grow on methanol. When one of these sequences was added to Plac of Escherichia coli, which is a weak constitutive promoter in M. extorquens AM1, the activity increased two- to threefold. These results suggest that this sequence is essential for normal expression of these genes in M. extorquens AM1, and may serve as a general enhancer element for genetic constructs in this bacterium.

scholarly journals Impacts of Gender Differences on Women’s Participation in Community Work in Abbay Chomman Woreda, Ethiopia

2019 ◽  
Vol 13 ◽  
pp. 86-118
Author(s):  
RATA B KENEA
Keyword(s):  
Economic Empowerment ◽  
Research Approach ◽  
Community Work ◽  
Quantitative And Qualitative Research ◽  
Women's Participation ◽  
Participation Of Women ◽  
The Status ◽  
The Difference ◽  
The Town ◽  
Status Of Women

Though the participation of women in economic development and community work usually realized low as a whole due to various reasons, the contribution of gender in creating the difference in terms of their status even among the participatory women is untouched area. For instance, the research conducted by Atinafu Diga (2013) on assessment of economic empowerment of women the related studies entitled Assessment of economic empowerment of Women in Kolobo kebele, Abay chomman Woreda dealt with only inadequate economic empowerment problems and low participation of Women in educational leadership areas respectively as their overall findings. As a result, this study is undertaken to assess the status of women disparity to participate in community work in case of kolobo kebele, Ambo town, Oromia Regional State; Ethiopia.This study is descriptive in nature. For this particular study, both quantitative and qualitative research approach were employed. In doing this research paper, of the total population,79 samples were selected from the town selected using simple random and purposive sampling techniques. Here, 79 were responded for the 20 close-ended questions of the questionnaire and another 10 were responded for open-ended questions of the interview. To analyses the quantitative data, techniques such as tabulation, percentages, and numerical figures were employed to. On the other hand, Narration and comparative discussion were covered to analyse the qualitatively collected data.

scholarly journals Territorial organization of the restaurant facilities of Kyiv agglomeration

2015 ◽  
pp. 79-82
Author(s):  
K. Dmytriieva
Keyword(s):  
Restaurant Industry ◽  
The Core ◽  
Industry Standards ◽  
The Status ◽  
Territorial Organization ◽  
The Difference ◽  
Surrounding Areas ◽  
Concentration Ratios

The article outlines the factors of development of the restaurant facilities in Kyiv and surrounding areas. The status of the territorial organization of the restaurant facilities of Kyiv and Kyiv region through the review of the main indicators of the industry by districts in 2013 has been analyzed. Comparison of the industry standards due to availability of these services to population needs has been performed. The result of the difference of efficiency of the restaurant industry through calculation of concentration ratios turnover has been received. Area that can be considered as the core of the restaurant industry of Kyiv agglomeration and area with the potential of growth has been identified. The article visualized by maps.

scholarly journals Reorientasi Gerakan Feminisme Islam; Sebuah Upaya Membangun Kesetaraan Perempuan

FIKROTUNA ◽  
2017 ◽  
Vol 3 (1) ◽  
Author(s):  
ABD WARITS
Keyword(s):  
Skin Color ◽  
Social Role ◽  
Human Beings ◽  
Men And Women ◽  
The World ◽  
The Status ◽  
History Of ◽  
The Difference ◽  
The Right ◽  
Arbitrary Action

In the history of women's life, the woman has never cracked from the wild cry of helplessness. Woman always become victim of men’s egoism, marginalized, hurt, unfettered, fooled and never appreciated the presence and role. This situation troubles many intellectual Muslims who have perspective that Islam teaches equality, equality for all human beings in the world. The difference in skin color, race, tribe and nation, as well as gender does not cause them to get the status of the different rights and obligations. The potential and the right to life of every human being and the obligation to serve the Lord Almighty is the same. Indeed, all human beings, as caliph in the world, have the same obligation, namely to prosperity of life in the world. No one is allowed to act arbitrarily, destroying, or hurt among others. They are required to live side by side, united, and harmonious, help each other and respect each other. However, that "demand" never becomes a reality. The differences among human identities become a barrier and the cause of divisions. For them, those who are outside environment, different identities are "others" who rightly do not need them "know". The difference of identity has become a reason to allow "hurt" each other. Several intellectual Muslims who recognize the wrong (discrimination against women), and then they attempt to formulate a movement for women's liberation. All the efforts have been done on the basis of awareness that arbitrary action by any person can never be justified. They also realize, that the backwardness of women are "stumbling block" that will lead to the resignation of a civilization. However, this struggle found a lot of challenges; including the consideration of "insubordination" to conquer the power of men, despite it had done by using many strategies. Starting from the writing of scientific book and countless fiction themed women has been published in order to give awareness of equality between men and women. This paper seeks to reexamine the process of the empowerment struggle to give a brand new concept, so that the struggle of women empowerment is not as insubordination and curiosity process in an attempt to conquer the male. Through approach of literature review and observations on the relationship between men and women, the writer finally concluded that the movement of Islamic feminism is not a movement to seize the power of men, but an attempt to liberate women from oppression so that they get the rights of their social role, giving freedom for women to pursue a career as wide as possible like a man, without forgetting a main duty as a mother: to conceive, give birth and breastfeed their children.

scholarly journals INTERNATIONAL LEGAL ORDER: ON THE ISSUE OF ITS PROPERTIES (THEORETICAL ASPECTS)

2016 ◽  
Vol 2 (127) ◽  
pp. 115-122
Author(s):  
I. Zabara
Keyword(s):  
International Law ◽  
The State ◽  
Legal Order ◽  
General Context ◽  
Theoretical Approaches ◽  
Ability To Act ◽  
The Status ◽  
International Legal Order ◽  
The Difference ◽  
Internal And External Factors

The article deals with one of the theoretical aspects of international legal order issues – the question of its properties. The author summarizes the doctrinal views of international law and regards the basic properties of the phenomenon of international legal order as its ability to act as system complexity, dynamism, orderliness, the reality and legitimacy of actions of subjects. The author notes that there is a common position in the doctrine, according to which the international legal order is a system. However, he notes that the difference in views on the international legal order as a system consists in the components the researchers include in its composition; the author examines two theoretical approaches. The complexity of the international legal order is determined from the standpoint of the number of its elements and components, as well as the number of their connections. This opinion highlights the fact that the predominant role is played by the quantity of links between elements and components, and indicates the international legal order capacity for permanent changes under the influence of the relevant internal and external factors. The dynamism of the international legal order is characterized from the point of capacity for the development and modification. It is stated that the state of the dynamics is effected by several circumstances. The author concludes that this international legal order’s property as a dynamism is one of the qualities that characterizes its condition as a system. The orderliness of the international legal order is considered from a consistency point, the interaction of parts of the whole, due to its structure. The author notes that the ordering of the international legal order displays its internal relationships and emphasizes its status as a system. The reality of the international legal order is characterized from the point of objectively existing phenomenon. The author concludes that the allocation of the international legal order of reality as one of its properties is intended to emphasize the status of one of its most important components - the state of international relations. Separately, the author considers the question of the legality of actions of subjects of international law, which are discussed in the doctrine from the standpoint of the conditions necessary for its maintenance. The author points out that in the general context of the properties that characterize the international legal order, it can be considered as an aspect wich together with other characterizes the state of the international legal order.

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