Data/hora: 01/06/2024 19:10:42

Provedor de dados: | Brazilian Journal of Microbiology
País: | Brazil
Título: | Cloning and high-level expression of Thermus thermophilus RecA in E. coli: purification and novel use in HBV diagnostics
Autores: | Sundarrajan,Sudarson; Rao,Sneha; Padmanabhan,Sriram.
Data: | 2018-12-01
Ano: | 2018
Palavras-chave: | RecA; Hepatitis B virus; DNA virus; Thermus thermophilus.
Resumo: | ABSTRACT We studied the role of Thermus thermophilus Recombinase A (RecA) in enhancing the PCR signals of DNA viruses such as Hepatitis B virus (HBV). The RecA gene of a thermophilic eubacterial strain, T. thermophilus, was cloned and hyperexpressed in Escherichia coli. The recombinant RecA protein was purified using a single heat treatment step without the use of any chromatography steps, and the purified protein (&gt;95%) was found to be active. The purified RecA could enhance the polymerase chain reaction (PCR) signals of HBV and improve the detection limit of the HBV diagnosis by real time PCR. The yield of recombinant RecA was ∼35 mg/L, the highest yield reported for a recombinant RecA to date. RecA can be successfully employed to enhance detection sensitivity for the diagnosis of DNA viruses such as HBV, and this methodology could be particularly useful for clinical samples with HBV viral loads of less than 10 IU/mL, which is interesting and novel.
Tipo: | Info:eu-repo/semantics/article
Idioma: | Inglês
Identificador: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822018000400848
Editor: | Sociedade Brasileira de Microbiologia
Relação: | 10.1016/j.bjm.2018.03.007
Formato: | text/html
Fonte: | Brazilian Journal of Microbiology  v.49 n.4 2018
Fonte: | info:eu-repo/semantics/openAccess