Pseudomonas is a communal motif of environmental associated disease and causes a serious problem in poultry farms, so this study was deliberated to investigate the quandary of Pseudomonas species especially Pseudomonas aeruginosa (P. aeruginosa) which has multifarious virulence genes and plays a major role in poultry outbreaks. Also, it focuses the light on the problem of antimicrobial and disinfectant resistance. A total of 200 samples (100 from dead in shell chicken embryos and 100 from broilers at different ages) were collected from different hatcheries and farms in Luxor governorate, 40 isolates (20%) of Pseudomonas species were isolated and identified serologically as P.cepacia, P.fluorescens, P.putida, P.fragi and P.aeruginosa. PCR inveterate the existence of P.aeruginosa DNA in seven isolates by using 16SrDNA primers at 956bp. P.aeruginosa isolates have different virulence genes such as toxA, exoS, lasB, lasI and oprL gene with incidence rate 71.42% for each of them, except oprL was 100%. Also, Quaternary Ammonium Compounds resistant genes (QACs) were detected in P. aeruginosa isolates with incidence rate (14.28%) for each of qacAB and qacCD genes, while the qacED1 gene incidence was (100%). P. aeruginosa isolates showed an obstacle of antimicrobial resistance for different antimicrobials while most of these isolates cleared susceptibility for ciprofloxacin and norfloxacin. In conclusion, this work described the problem of P. aeruginosa as it proved a high virulence repertoire owned by the P. aeruginosa that confirming its pathogenicity for chicken embryos and broilers. Also, our study is fuelling the concern on disinfectant resistance problem and displaying the relation between QACs and antibiotic resistance. So, the deterrence of the Pseudomonas infection in the poultry housing becomes necessary by applying strict bio-security measures.

DNA regions that flank a gene's promoter play an important role in determining transcription efficiency by interacting with the carboxy-terminal domain of RNA polymerase alpha-subunit. We placed an adenine-rich upstream element (UP) between -38 and -59 of the core trc promoter to enhance gene expression in Ochrobactrum anthropi up to 66-fold. The high level of expression achieved by the UP element and the N-terminus fusion of a 6xHis epitope tag facilitated detection and purification of heterologous proteins directly from O. anthropi.

The creation of a double His-tag fusion that forms a RNA stem loop in the mRNA encoding the N-terminus of the target protein is a novel approach for the enhancement of expression, purification, and detection of a recombinant protein. Compared to a single His-tag fusion, a tandem His-tag fusion RNA stem loop, located downstream of the constitutive groE and Ch promoters, enhanced heterologous gene expression in Brucella, Salmonella, and Escherichia. We demonstrated one-step detection and purification of recombinant green fluorescence protein (GFP) directly from Brucella spp. without using Escherichia coli as an expression host. The amount of purified GFP using the tandem His-tag RNA stem loop increased more than threefold; moreover, the sensitivity of detection increased more than fourfold in comparison to the single His-tag fusion form. This method has the potential to significantly improve heterologous gene expression and high-throughput protein synthesis and purification.