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Dergiler / Turkish Journal of Biology / 2015 / Cilt: 39 - Sayı: 2

Prokaryotic expression, purifcation, polyclonal antibody preparation, and tissue distribution of porcine Six1

Wanxue WEN
Shuai CHANG
Xiaoyan WANG
Junqiu LUO
Meng XU
Xiaoling CHEN
Zhiqing HUANG
Guangmang LIU
Daiwen CHEN
Bing YU
Sayfa
335–342
Yayın Tarihi
2022-11-05
DOI
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⚡ TL;DR(1 Cümlede Hızlı Çıkarım)

The tissue distribution of pSix1 was determined by western blot using the prepared polyclonal antibody.

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Amaç & Hipotez:Sine oculis homeobox 1 (Six1), a member of the Six homeoproteins, plays an important role in skeletal myogenesis and thespecifcation of myofber diversity.
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Temel Bulgular & Veri:In this study, in order to scale up the production of recombinant porcine Six1 (pSix1), a pET-30a(+)-pSix1 plasmid was constructed and transformed into Escherichia coli BL21 (DE3).
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Sonuç & Katkı:The tissue distribution of pSix1 was determined by western blot using the prepared polyclonal antibody.
🔬Tür:Araştırma Makalesi
📍Kapsam:Genişletilmiş Akademik Örneklem
⚙️Değişkenler:Alan Parametreleri

Abstract

Sine oculis homeobox 1 (Six1), a member of the Six homeoproteins, plays an important role in skeletal myogenesis and thespecifcation of myofber diversity. In this study, in order to scale up the production of recombinant porcine Six1 (pSix1), a pET-30a(+)-pSix1 plasmid was constructed and transformed into Escherichia coli BL21 (DE3). The recombinant pSix1 could be induced for efcientexpression with 2 mM IPTG for 2 h at 30 °C, yielding approximately 4.6 mg/L. The protein was then purifed and identifed by westernblot, and it was used for preparing its polyclonal antibody. The recombinant pSix1 was tagged with a 6-His tag at its C-terminus, whichcould be conveniently purifed by afnity column. The purifed recombinant protein was used for immunizing Sprague Dawley rats toobtain the polyclonal antibody against pSix1. The antibody titer and specifcity were determined by ELISA and western blot analysis,respectively. The tissue distribution of pSix1 was determined by western blot using the prepared polyclonal antibody.

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Bu Makaleye Atıf Yap

WEN, W., CHANG, S., WANG, X., LUO, J., XU, M., CHEN, X., HUANG, Z., LIU, G., CHEN, D., YU, B. (2022). Prokaryotic expression, purifcation, polyclonal antibody preparation, and tissue distribution of porcine Six1. Turkish Journal of Biology, Cilt: 39 - Sayı: 2, 335–342. https://ajindex.com/articles/654430

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