کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
2079056 | 1545067 | 2006 | 7 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Preparation of Recombinant HIV-TATm-Survivin (T34A) Protein and Its Proapoptosis Activity to Four Cancer Cell Lines in vitro
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کلمات کلیدی
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
بیوتکنولوژی یا زیستفناوری
پیش نمایش صفحه اول مقاله
![عکس صفحه اول مقاله: Preparation of Recombinant HIV-TATm-Survivin (T34A) Protein and Its Proapoptosis Activity to Four Cancer Cell Lines in vitro Preparation of Recombinant HIV-TATm-Survivin (T34A) Protein and Its Proapoptosis Activity to Four Cancer Cell Lines in vitro](/preview/png/2079056.png)
چکیده انگلیسی
In this study, the cDNA encoding survivin was cloned by RT-PCR obtained from human breast cancer cell lines, B-Cap37. After the mutation of survivin (T34A) by site-directed mutagenesis, an expression vector of pRSET-B-HIV-TATm-Survivin (T34A) was constructed by PCR. Subsequently, the resultant plasmid was transformed into E. coli BL21 (DE3). The recombinant HIV-TATm-Survivin (T34A) protein was expressed efficiently by inducing IPTG at a concentration of 0.5 mmol/L, reaching a yield of 650 mg/L (as inclusion body) in fermentation culture. The inclusion bodies were solubilized, refolded, and purified by ion exchange chromatography and size-exclusion chromatography to a purity of 96 %. Remarkable effects of the purified recombinant HIV- TATm-Survivin (T34A) on the morphology of cell line SW1990 and B-Cap37 were observed after being administrated for 4 h. MTT assay showed that the recombinant HIV-TATm-Survivin (T34A) protein could restrain the cell proliferation of SW1990, B-Cap37, and SSMC-7721, in vitro. Apoptosis rate and cell circle of SW1990 and B-Cap37 that were treated with the target protein (final concentration 60 μg/mL) were detected with flow cytometry. The results revealed that the apoptosis rate of SW1990 and B-Cap37 were 25.6 % and 19.3 %, respectively. More than 65 % of cancer cells were arrested at the G1 phase. The study suggested that TATm-Survivin (T34A) protein was a hopeful protein drug for the treatment of cancer by facilitating apoptosis of cancer cells.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Chinese Journal of Biotechnology - Volume 22, Issue 2, March 2006, Pages 285-292
Journal: Chinese Journal of Biotechnology - Volume 22, Issue 2, March 2006, Pages 285-292
نویسندگان
ZHENG Wen-Yun, MA Xing-Yuan, WEI Dong-Zhi, WANG Jin-Zhi, LIU Qing-Hai, MA Yu-Shu, YANG Sheng-Li,