Using overlap ‐extension PCR technique to fusing genes for constructing recombinant plasmids
Overlap‐extension PCR is a method for splice of gene segments to produce focused fragments for constructing recombinant plasmid, but its complexity limits its application. To simplify the protocol and to improve the effectiveness, we employed gradient temperatures to replace the single annealing temperature in the thermo‐cycling program, and optimize the templates ratio. The concentration of each fragment was adjusted to 10 ng µl−1. Fragment concentration ratio was the inverse of the fragment size ratio. The products of fused segments were 2000–5000 bp in length using the revised one‐step method. This meth...
Source: Journal of Basic Microbiology - December 1, 2017 Category: Microbiology Authors: Yuanyuan Lu, Shuqin Xiao, Mingyue Yuan, Yibo Gao, Jiaying Sun, Chunsheng Xue Tags: METHOD PAPER Source Type: research
Autolytic enzymes are responsible for increased melanization of carbon stressed Aspergillus nidulans cultures
Melanization of carbon stressed Aspergillus nidulans cultures were studied. Melanin production showed strong positive correlation with the activity of the secreted chitinase and ß‐1,3‐glucanase. Deletion of either chiB encoding an autolytic endochitinase or engA encoding an autolytic ß‐1,3‐endoglucanase, or both, almost completely prevented melanization of carbon stressed cultures. In contrast, addition of Trichoderma lyticase to cultures induced melanin production. Synthetic melanin could efficiently inhibit the purified ChiB chitinase activity. It could also efficiently decrease the intensity of hyphal fragment...
Source: Journal of Basic Microbiology - December 1, 2017 Category: Microbiology Authors: Melinda Szil ágyi, Fruzsina Anton, István Pócsi, Tamás Emri Tags: RESEARCH PAPER Source Type: research
A minireplicon of plasmid pBMB26 represents a new typical replicon in the megaplasmids of Bacillus cereus group
A new minireplicon (rep26 minireplicon) from pBMB26, the 188 kb indigenous plasmid related to spore‐crystal association (SCA) phenotype in Bacillus thuringiensis strain YBT‐020, was characterized. A 12 kb EcoRI fragment, which encoded 10 putative open reading frames (ORFs), was capable of supporting replication when cloned in a replication probe vector. Deletion and frame shift mutation analysis showed that a 4.1 kb region encompassing two putative ORFs (orf21 and orf22) was essential for the plasmid replication in B. thuringiensis. Gene orf21 encoding a 49.8 kDa protein (named Rep26) with a helix‐turn‐heli...
Source: Journal of Basic Microbiology - December 1, 2017 Category: Microbiology Authors: Pengxia Wang, Yiguang Zhu, Hui Shang, Yun Deng, Ming Sun Tags: RESEARCH PAPER Source Type: research
Metal ‐citrate complex transport in Kineococcus radiotolerans
The growth of an organism is highly dependent on the acquisition of carbon and metals, and availability of these nutrients in the environment affects its survival. Organisms can obtain both nutrients simultaneously through proteins of the CitMHS superfamily. Bioinformatic studies suggested a CitMHS gene (Accession number ABS03965.1) in Kineococcus radiotolerans. Radio flux assays following 14‐C radiolabelled citrate, either free or complexed to a variety of metal ions, in K. radiotolerans demonstrated internalization of the citrate when bound to select metal ions only, primarily in the form of calcium‐citrate. A pH res...
Source: Journal of Basic Microbiology - December 1, 2017 Category: Microbiology Authors: Brian P. Huta, Nigel H. Miller, Eleanor L. Robertson, Robert P. Doyle Tags: RESEARCH PAPER Source Type: research
Identification of virus ‐derived siRNAs and their targets in RBSDV‐infected rice by deep sequencing
We report for the first time vsiRNAs derived from RBSDV‐infected rice. (Source: Journal of Basic Microbiology)
Source: Journal of Basic Microbiology - December 1, 2017 Category: Microbiology Authors: Ying Lan, Yanwu Li, Zhiguo E, Feng Sun, Linlin Du, Qiufang Xu, Tong Zhou, Yijun Zhou, Yongjian Fan Tags: RESEARCH PAPER Source Type: research
Transcriptional regulation of acetyl CoA and lipid synthesis by PII protein in Synechococcus PCC 7942
In this study, impacts of PII deficiency on the synthesis of acetyl CoA and acetyl CoA carboxylase enzyme (ACCase) was analyzed in the Synechococcus sp. PCC 7942 by evaluating the mRNA levels of pyruvate kinase (PK), pyruvate dehydrogenase (PDH), citrate synthase (CS), biotin synthase (BS), biotin carboxylase (BC), biotin carboxyl carrier protein (BCCP), carboxyl transferase (CT) α and β subunits. The PII deficient Synechococcus sp. PCC 7942 showed upregulation of all the above‐mentioned genes, except CS. Analyses of genes required for acetyl coA synthesis exhibited a substantial increase in the transcript levels of PK...
Source: Journal of Basic Microbiology - December 1, 2017 Category: Microbiology Authors: Ekta Verma, Sindhunath Chakraborty, Balkrishna Tiwari, Arun K. Mishra Tags: RESEARCH PAPER Source Type: research
Production, purification, and characterization of a novel serine ‐esterase from Aspergillus westerdijkiae
Esterases hydrolyze water soluble short chain fatty acids esters and are biotechnologically important. A strain of Aspergillus westerdijkiae isolated from cooking oil for recycling was found to secrete an esterase. The best enzyme production (19‐24 U/ml of filtrate) culture conditions were stablished. The protein was purified using ammonium sulphate precipitation, dialysis, and a chromatographic step in Sephacryl S‐200 HR. The 32 kDa purified protein presented an optimal temperature of 40°C, with a T50 of 48.95°C, and an optimal pH of 8.0. KM and Vmax were 638.11 µM for p‐NPB and 5.47 µmol of released p...
Source: Journal of Basic Microbiology - November 28, 2017 Category: Microbiology Authors: Fausto F. Castro, Ana B. P. Pinheiro, Edileusa C. M. Gerhardt, Marco A. S. Oliveira, Ione P. Barbosa ‐Tessmann Tags: RESEARCH PAPER Source Type: research
Biosynthetic energy cost of potentially highly expressed proteins vary with niche in selected actinobacteria
Amino acid and protein biosynthesis requires a number of high energy phosphate bonds and includes a dual energy cost for the synthesis of chemical intermediates during the fueling reactions and the conversion of precursor molecules to final products. One popular hypothesis is that the proteins encoded by putative highly expressed genes (hence called PHXPs) generally utilize low energy consuming amino acids to reduce the biosynthetic cost of the essential proteins. In our study, we found that this idea was not supported in the case of actinobacteria. With the actinobacteria, the energy costs of PHXPs varied in relation to t...
Source: Journal of Basic Microbiology - November 16, 2017 Category: Microbiology Authors: Indrani Sarkar, Louis S. Tisa, Maher Gtari, Arnab Sen Tags: RESEARCH PAPER Source Type: research
Statistically optimized ceftriaxone sodium biotransformation through Achromobacter xylosoxidans strain Cef6: an unusual insight for bioremediation
The present study underlines a unique promising approach toward efficient biotransformation of ceftriaxone sodium (Ceftx), a highly frequent prescribed cephalosporin antibiotic, by a newly bacterium namely Achromobacter xylosoxidans strain Cef6 isolated from Ceftx contaminated raw materials in pharmaceutical industries. A three step sequential statistical‐mathematical approach (Plackett‐Burman design [PBD], Central Composite Design [CCD], and ridge‐canonical analyses) was anticipated to optimize the biotransformation process. Ceftx concentration and medium volume: bottle volume ratio, two key determinants, significan...
Source: Journal of Basic Microbiology - November 15, 2017 Category: Microbiology Authors: Ahmed Anan, Khaled M. Ghanem, Amira M. Embaby, Ahmed Hussein, Moustafa Y. El ‐Naggar Tags: RESEARCH PAPER Source Type: research
Morphological and cellular diversity of magnetotactic bacteria: A review
Journal of Basic Microbiology, EarlyView. (Source: Journal of Basic Microbiology)
Source: Journal of Basic Microbiology - November 7, 2017 Category: Microbiology Source Type: research
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Journal of Basic Microbiology, Ahead of Print. (Source: Journal of Basic Microbiology)
Source: Journal of Basic Microbiology - November 7, 2017 Category: Microbiology Source Type: research
Issue Information: Journal of Basic Microbiology. 11/2017
(Source: Journal of Basic Microbiology)
Source: Journal of Basic Microbiology - November 7, 2017 Category: Microbiology Tags: ISSUE INFORMATION Source Type: research
Cover: Journal of Basic Microbiology. 11/2017
Cover illustration:
Fungi are able to use xylose, shown here for anaerobic fungi co‐cultured with methanogens; formate and hydrogen were utilized by the associated methanogens. The associated methanogens intensified the metabolism in hydrogenosomes of anaerobic fungi with xylose as substrate (for abbreviations, see pp. 933–940, this issue).
Background hyphae (Photo: Erika Kothe, Friedrich‐Schiller‐University, Jena, Germany) (Source: Journal of Basic Microbiology)
Source: Journal of Basic Microbiology - November 7, 2017 Category: Microbiology Tags: COVER PICTURE Source Type: research
CntA oxygenase substrate profile comparison and oxygen dependency of TMA production in Providencia rettgeri
CntA oxygenase is a Rieske 2S‐2Fe cluster‐containing protein that has been previously described as able to produce trimethylamine (TMA) from carnitine, gamma‐butyrobetaine, glycine betaine, and in one case, choline. TMA found in humans is exclusively of bacterial origin, and its metabolite, trimethylamine oxide (TMAO), has been associated with atherosclerosis and heart and renal failure. We isolated four different Rieske oxygenases and determined that there are no significant differences in their substrate panels. All three had high activity toward carnitine/gamma‐butyrobetaine, medium activity toward glycine betai...
Source: Journal of Basic Microbiology - November 7, 2017 Category: Microbiology Authors: Gints Kalnins, Eduards Sevostjanovs, Dace Hartmane, Solveiga Grinberga, Kaspars Tars Tags: RESEARCH PAPER Source Type: research
Expression, purification, and characterization of a novel laccase from Setosphaeria turcica in Eschericha coli
In this study, a novel laccase, Stlac4, from Setosphaeria turcica was cloned and expressed in Escherichia coli by insertion into the pET‐30a expression plasmid. The recombinant laccase was purified and visualized on SDS–PAGE as a single band with an apparent molecular weight of 71.5 KDa, and confirmed by Western blot. The maximum activity of the purified laccase was 127.78 U · mg−1, the optimum temperature and pH value were 60 °C and 4.0 respectively, measured by oxidation of 2,2′‐Azinobis‐(3‐ethylbenzthiazoline‐6‐sulphonate) (ABTS). Purified laccase activity under different metal ions and an in...
Source: Journal of Basic Microbiology - November 7, 2017 Category: Microbiology Authors: Shuangxin Ma, Ning Liu, Hui Jia, Dongqing Dai, Jinping Zang, Zhiyan Cao, Jingao Dong Tags: RESEARCH PAPER Source Type: research
