Expression and Characterization of the Staphylococcus aureus RecA protein: A mapping of canonical functions

2022 ◽  
Vol 189 ◽  
pp. 105967
Author(s):  
Kajal Kiran ◽  
K. Neelakanteshwar Patil
Keyword(s):  
Staphylococcus Aureus ◽  
Protein A ◽  
Reca Protein

scholarly journals Whole-Genome Epidemiology and Characterization of Methicillin-Susceptible Staphylococcus aureus ST398 From Retail Pork and Bulk Tank Milk in Shandong, China

2021 ◽  
Vol 12 ◽  
Author(s):  
Xiaonan Zhao ◽  
Ming Hu ◽  
Cui Zhao ◽  
Qing Zhang ◽  
Lulu Li ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Protein A ◽  
Whole Genome ◽  
Staphylococcal Protein A ◽  
Sequencing Analysis ◽  
Bulk Tank Milk ◽  
Predominant Type ◽  
Bulk Tank ◽  
The One

Staphylococcus aureus (S. aureus) is now regarded as a zoonotic agent. Methicillin-susceptible S. aureus (MSSA) ST398 is a livestock-associated bacterium that is most prevalent in China, but there are currently no data available for Shandong. Therefore, the aim of this study was to investigate the epidemiology and characterization of MSSA ST398 from retail pork and bulk tank milk (BTM) in Shandong. A total of 67 S. aureus isolates were collected from retail pork between November 2017 and June 2018. Among the isolates, high antimicrobial resistance rates were observed for penicillin (97.0%), and 92.5% of the isolates were multi-drug resistant (MDR). Eight sequence types (STs) were identified in the retail pork isolates, and the predominant type was ST15 (n=26), which was followed by ST398 (n=14). Staphylococcal protein A gene (spa) typing identified spa types t034 and t1255 in MSSA ST398 from retail pork. Using whole-genome sequencing analysis, we described the phylogeny of 29 MSSA ST398 isolates that were obtained from retail pork (n=14) and BTM (n=15). The phylogenetic tree showed that the MSSA ST398 isolates from different sources had the same lineage. Among the 29 MSSA ST398 isolates, five resistance genes were detected, and all isolates carried DHA-1. Fifteen toxin genes were detected, and all isolates carried eta, hla, and hlb. In conclusion, this study found that a high risk for MSSA ST398 was present in retail pork and BTM. These findings have major implications for how investigations of MSSA ST398 outbreaks should be conducted in the One-Health context.

scholarly journals Characterization of NorR Protein, a Multifunctional Regulator of norA Expression in Staphylococcus aureus

2003 ◽  
Vol 185 (10) ◽  
pp. 3127-3138 ◽  
Author(s):  
Que Chi Truong-Bolduc ◽  
Xiamei Zhang ◽  
David C. Hooper
Keyword(s):  
Gene Expression ◽  
Staphylococcus Aureus ◽  
Efflux Pump ◽  
Protein A ◽  
Protein Overexpression ◽  
Wild Type ◽  
Liquid Media ◽  
Cell Surface Properties ◽  
Regulatory Systems

ABSTRACT We characterized a Staphylococcus aureus norA gene expression regulator, NorR, initially identified from its binding to the norA promoter. The norR gene was 444 bp in length, located ∼7 kb upstream from the norA gene, and encoded a predicted 17.6-kDa protein. Overexpression of norR in wild-type S. aureus strain ISP794 led to a fourfold decrease in sensitivity to quinolones and ethidium bromide and an increase in the level of norA transcripts, suggesting that NorR acts as a positive regulator of norA expression. Overexpression of norR in sarA and agr mutants did not alter quinolone sensitivity or levels of norA transcription, indicating that the presence of these two global regulatory systems is necessary for NorR to affect the expression of norA. Insertion and disruption of norR in ISP794 increased resistance to quinolones by 4- to 16-fold but had no effect on norA transcription, suggesting that NorR acts as a repressor for another unidentified efflux pump or pumps. These mutants also exhibited an exaggerated clumping phenotype in liquid media, which was complemented fully by a plasmid-encoded norR gene. Collectively, these results indicate that NorR is a multifunctional regulator, affecting cell surface properties as well as the expression of NorA and likely other multidrug resistance efflux pumps.

scholarly journals Isolation from Sewage Influent and Characterization of Novel Staphylococcus aureus Bacteriophages with Wide Host Ranges and Potent Lytic Capabilities

2009 ◽  
Vol 75 (13) ◽  
pp. 4483-4490 ◽  
Author(s):  
Aidan J. Synnott ◽  
Ying Kuang ◽  
Miki Kurimoto ◽  
Keiko Yamamichi ◽  
Hidetomo Iwano ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Host Range ◽  
Protein A ◽  
Bovine Mastitis ◽  
Economic Consequences ◽  
Resistant Bacteria ◽  
Bacteriophage Therapy ◽  
Transmission Electron ◽  
65 H

ABSTRACT Staphylococcus aureus is a gram-positive pathogen that causes a variety of diseases, including bovine mastitis, which has severe economic consequences. Standard antibiotic treatment results in selection of resistant strains, leading to a need for alternative treatments, such as bacteriophage therapy. Forty-nine S. aureus isolates were obtained from the milk of mastitic cows for use in screening of staphylococcal phages. Fifteen isolates which were positive for both coagulase and hemolysin were assayed by PCR for variation in the X region and the immunoglobulin G-binding region of the protein A gene (spa) and in the carboxy terminus of the coagulase gene (coa) and for the presence of enterotoxin C, G, H, and I genes. The host ranges of 52 phages isolated from sewage influent were determined by performing spot tests with the 15 S. aureus isolates, and two phages were subsequently chosen for further analysis. ΦSA039 had the widest host range, producing clear plaques on 13 of the 15 isolates (87%), while ΦSA012 produced clear plaques on 8 isolates (53%) and was the only phage that produced a clear plaque on a nonmastitic S. aureus strain. Transmission electron microscopy revealed that the phages were similar sizes and belonged to the Myoviridae family. Measurement of optical densities during coculture with S. aureus isolates confirmed the breadth of the ΦSA039 host range and showed that ΦSA012 had potent lytic capability. ΦSA012-resistant bacteria did not appear for three of seven isolates tested (43%) after 65 h of incubation. These two phages are proposed as candidates for phage therapy of bovine mastitis.

Bacteriological and Molecular Identification and Characterization of Staphylococcus aureus from Different Affections of Canines

2020 ◽  
Author(s):  
Sejal S. Chaudhary ◽  
Harshad C. Chauhan ◽  
Kishan Kumar Sharma ◽  
Sandip S. Patel ◽  
Sushil Kumar Mohapatra ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Protein A ◽  
Molecular Techniques ◽  
Haemolytic Activity ◽  
Eye Infections ◽  
Respiratory Problems ◽  
Spa Gene ◽  
Staphylococcus Spp ◽  
Identification And Characterization

Background: The present study was done to ascertain prevalence of Staphylococcus aureus from various canine affections in Banaskantha district of Gujarat, India. Along with this, use of classical and molecular techniques were compared in identification and virulence characterization of this pathogen.Methods: A total of 165 samples were collected and bacterial identification was carried out with bacteriological (phenotypic) techniques and confirmed by genus specific 16S rDNA and Staphylococcus aureus specific sa442 gene based PCR. Isolates were characterized for coagulase production, haemolysis activity and presence of spa gene. Result: Samples yielded, 88 (53.33%) Staphylococcus spp. via bacteriological and PCR methods. Clinically, 19 (21.59%), 28 (31.82%), 12 (13.64%), 15 (17.04%) and 14 (15.91%) isolates were from abscess/wound, pyoderma, respiratory problems, eye infections and Otitis, respectively. A total of 46/88 (52.27%) isolates were confirmed as Staphylococcus aureus in PCR. In tube coagulase test, 51/88 (57.95%) isolates were found positive. A tota of 42 isolates revealed presence of coa gene, including two tube coagulase negative isolates. Haemolytic activity revealed beta (51.14%) gamma (31.82%), alpha (13.64%) and alpha-beta (3.41%) haemolysis, respectively. X-region of Protein A (spa gene) was detected in 26 /46 (56.52%) isolates in PCR.

scholarly journals Epidemiological characterization of methicillin-resistant Staphylococcus aureus isolated in the North West of England by protein A (spa) and coagulase (coa) gene polymorphisms

1998 ◽  
Vol 121 (3) ◽  
pp. 507-514 ◽  
Author(s):  
J. WALKER ◽  
R. BORROW ◽  
V. EDWARDS-JONES ◽  
B. A. OPPENHEIM ◽  
A. J. FOX
Keyword(s):  
Staphylococcus Aureus ◽  
Gene Polymorphisms ◽  
Protein A ◽  
Pcr Amplification ◽  
Strain Identification ◽  
Methicillin Resistant ◽  
North West ◽  
The North ◽  
Length Polymorphisms

In a comparative study, isolates of methicillin-resistant Staphylococcus aureus (MRSA) with known pulsed-field gel electrophoresis (PFGE) and bacteriophage type were analysed by polymerase chain reaction (PCR) and restriction fragment length polymorphisms (RFLP) for additional discriminatory subtyping information. PFGE was previously performed using standardized, commercially available kits and pre-programmed software. Isolates were examined for coagulase (coa) and protein A (spa) gene polymorphisms following PCR amplification of the coa hypervariable and spa repeat regions. Coa gene RFLPs produced a total of 38 distinct combined patterns after digestion with HaeIII and AluI and identified the predominant epidemic (EMRSA) types 15 and 16. A unique HaeIII restriction site was identified by RFLP and sequence analysis in the coa gene for EMRSA 15 but not EMRSA 16. The spa gene PCR yielded a total of 14 different profiles ranging from 3–18 repeats with the 2 predominant EMRSA types falling into 2 distinct groups. PCR detection of coa and spa polymorphisms offer a rapid preliminary strain identification and discriminatory subtyping information for surveillance of MRSA.

Molecular characterization of Staphylococcus aureus in Lebanon

2010 ◽  
Vol 138 (5) ◽  
pp. 707-712 ◽  
Author(s):  
S. T. TOKAJIAN ◽  
P. ABOU KHALIL ◽  
D. JABBOUR ◽  
M. RIZK ◽  
M. J. FARAH ◽  
...  
Keyword(s):  
Public Health ◽  
Staphylococcus Aureus ◽  
Molecular Characterization ◽  
Protein A ◽  
Multi Locus Sequence Typing ◽  
Health Concern ◽  
Human Pathogen ◽  
Public Health Concern ◽  
Sequence Types

SUMMARYStaphylococcus aureus is an important human pathogen and is a growing public health concern. In this study, 130 S. aureus, 93 methicillin-resistant S. aureus (MRSA) and 37 methicillin-sensitive S. aureus (MSSA), clinical isolates recovered from Lebanon were typed by protein A gene (spa) sequencing and multi-locus sequence typing (MLST). Forty-eight different spa types were identified and clustered into 30 different groups. MLST revealed 10 sequence types (STs) among the isolates. There were eight major MRSA clones defined as isolates with the same ST and the same SCCmec type. The majority of the PVL-positive isolates (53%) were ST80-MRSA-IVc. Systematic surveillance of both hospital and community isolates in Lebanon together with measures designed to limit the spread are required.

Comparative sequence analysis of spa gene of Staphylococcus aureus isolated from bovine mastitis: characterization of an unusual spa gene variant

2006 ◽  
Vol 73 (3) ◽  
pp. 322-327 ◽  
Author(s):  
Amr El-Sayed ◽  
Jörg Alber ◽  
Christoph Lämmler ◽  
Amir Abdulmawjood ◽  
Michael Zschöck ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Protein A ◽  
Bovine Mastitis ◽  
Clinical Mastitis ◽  
Comparative Sequence Analysis ◽  
Border Region ◽  
Binding Domains ◽  
Igg Binding ◽  
Spa Gene

The protein A encoding gene spa of four Staphylococcus aureus strains isolated from bovine clinical mastitis was amplified by PCR and sequenced. The four strains were selected after an initial screening of spa gene of 41 strains isolated from mastitic cows and were subjected to detailed investigations. According to the sequencing results the spa gene of three strains (M1, M2, M3) appeared with gene segments encoding five (E, D, A, B and C) and four (E, A, B and C) IgG binding domains for two (M1, M3) and one (M2) strain, respectively and with gene segments encoding four, two and two repeats of the octapeptide Xr-repeats for the strains M1, M2 and M3, respectively. For the remaining Staph. aureus strain (M4) gene segments encoding IgG binding domains E, D and A and a new domain BC with a size of 219 bp could be observed. The BC domain appears, with a deletion of a 123 bp segment from the border region between both domains, as fused domain of both previously characterized domains. The Xr-region of this strain had 11 octapeptide repeats.

Characterization of a Tn551-mutant of Staphylococcus aureus defective in the production of several exoproteins

1994 ◽  
Vol 40 (8) ◽  
pp. 677-681 ◽  
Author(s):  
Ana T. Giraudo ◽  
Claudia G. Raspanti ◽  
Aldo Calzolari ◽  
Rosa Nagel
Keyword(s):  
Staphylococcus Aureus ◽  
Regulatory Gene ◽  
Blot Hybridization ◽  
Protein A ◽  
Southern Blot Hybridization ◽  
Terminal Segment ◽  
Transcriptional Level ◽  
Extracellular Form ◽  
Single Insertion

A Tn551 insertional pleiotropic mutant defective in the production of several exoproteins was isolated from Staphylococcus aureus 196E and characterized. The pleiotropism of the mutant was due to a single insertion of the transposon as evidenced by Southern blot hybridization and by the transfer of its phenotype by transduction to S. aureus ISP479. The mutants showed diminished or null levels of α- and β-hemolysins, DNase, coagulase, and protein A in the supernatants of broth cultures. Production of proteases, lipase, staphylokinase, or enterotoxin A was not modified. The mutants did synthesize the cell-bound form of protein A and also the extracellular form of this protein coded by pRIT11, which lacks the COOH-terminal segment of the molecule. These observations suggest that the sae locus does not involve a positive regulatory gene acting at the transcriptional level. The phenotype of the mutant was different from that of other insertional mutants affecting exoprotein synthesis, such as agr, xpr, or sar. This new mutation has been designated sae (for S. aureus exoprotein expression).Key words: S. aureus, transpositional mutant, exoprotein production, pleiotropism, protein A.

Phylogeny Characterization of Seb Gene Encoding Enterotoxins in Staphylococcus aureus Isolated from Raw Milk and Cheese

2019 ◽  
Vol 9 (o3) ◽  
Author(s):  
Fatima N. Aziz ◽  
Laith Abdul Hassan Mohammed-Jawad
Keyword(s):  
Staphylococcus Aureus ◽  
Food Poisoning ◽  
Raw Milk ◽  
Staphylococcal Enterotoxin B ◽  
Human Pathogen ◽  
Conventional Pcr ◽  
Biochemical Tests ◽  
Specific Primers ◽  
Gene Encoding

Food poisoning due to the bacteria is a big global problem in economically and human's health. This problem refers to an illness which is due to infection or the toxin exists in nature and the food that use. Milk is considered a nutritious food because it contains proteins and vitamins. The aim of this study is to detect and phylogeny characterization of staphylococcal enterotoxin B gene (Seb). A total of 200 milk and cheese samples were screened. One hundred ten isolates of Staphylococcus aureus pre-confirmed using selective and differential media with biochemical tests. Genomic DNA was extracted from the isolates and the SEB gene detects using conventional PCR with specific primers. Three staphylococcus aureus isolates were found to be positive for Seb gene using PCR and confirmed by sequencing. Sequence homology showed variety range of identity starting from (100% to 38%). Phylogenetic tree analyses show that samples (6 and 5) are correlated with S. epidermidis. This study discovered that isolates (A6-RLQ and A5-RLQ) are significantly clustered in a group with non- human pathogen Staphylococcus agnetis.

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