کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
11013169 | 1797282 | 2018 | 8 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Cloning and high-level expression of Thermus thermophilus RecA in E. coli: purification and novel use in HBV diagnostics
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موضوعات مرتبط
علوم زیستی و بیوفناوری
ایمنی شناسی و میکروب شناسی
میکروبیولوژی و بیوتکنولوژی کاربردی
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چکیده انگلیسی
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 (>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 â¼35Â 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 10Â IU/mL, which is interesting and novel.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Brazilian Journal of Microbiology - Volume 49, Issue 4, OctoberâDecember 2018, Pages 848-855
Journal: Brazilian Journal of Microbiology - Volume 49, Issue 4, OctoberâDecember 2018, Pages 848-855
نویسندگان
Sudarson Sundarrajan, Sneha Rao, Sriram Padmanabhan,