Difference between revisions of "Part:BBa K1405002"

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−	<a title="Figure 2 Native-PAGE to provide the molybdate lane 1, the modA(acetic buffer PH 5.0); lane 2, the modA incubate with 20mM molybdate." href="https://static.igem.org/mediawiki/2014/9/90/Bnu_moda4.jpg" rel="prettyPhoto"> <span class="overlay zoom" style="display: none;"></span><img width="60%" style="margin-left:180px" src="https://static.igem.org/mediawiki/2014/e/e5/Bnu_moda4_01.jpg"> </a>	+	<a title="Figure 2 Native-PAGE to provide the molybdate lane 1, the modA(acetic buffer PH 5.0); lane 2, the modA incubate with 20mM molybdate." href="https://static.igem.org/mediawiki/2014/9/90/Bnu_moda4.jpg" rel="prettyPhoto"> <span class="overlay zoom" style="display: none;"></span><img width="45%" style="margin-left:250px" src="https://static.igem.org/mediawiki/2014/e/e5/Bnu_moda4_01.jpg"> </a>
	<p class="fig" style="width:85%; margin-left:50px">Figure 2 Native-PAGE to provide the molybdate lane 1, the modA(acetic buffer PH 5.0); lane 2, the modA incubate with 20mM molybdate.</p>		<p class="fig" style="width:85%; margin-left:50px">Figure 2 Native-PAGE to provide the molybdate lane 1, the modA(acetic buffer PH 5.0); lane 2, the modA incubate with 20mM molybdate.</p>
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Revision as of 14:13, 17 October 2014

Background

modA a periplasmic binding protein, . had a affinity (Km) for molybdate of 3–5 mM, this is one of the highest substrate Kd values reported for any periplasmic binding protein, the BBa_K1405002 contains the ModA coding sequence. And we used pET-21a BL21 to express and use His-chelating chromatography to purify.

Result

Figure 1 Expression and purification of ModA. Expression of ModA and its purification by His-chelating chromatography. Lane 1, molecular weight standards (kDa); lane 2, total bacterial proteins before IPTG induction; lane 3, total bacterial proteins after 0.5 mM IPTG induction; lane 4, total bacterial lysate after 0.5 mM IPTG induction; lane 5,the sediment after 16,000 rpm 30 min; lane 6 the flow through of column loading; lane 7,the flow of buffer B .lane 8 The purification of ModA by size-elution chromatography.

Figure 1 Expression and purification of ModA. Expression of ModA and its purification by His-chelating chromatography. Lane 1, molecular weight standards (kDa); lane 2, total bacterial proteins before IPTG induction; lane 3, total bacterial proteins after 0.5 mM IPTG induction; lane 4, total bacterial lysate after 0.5 mM IPTG induction; lane 5,the sediment after 16,000 rpm 30 min; lane 6 the flow through of column loading; lane 7,the flow of buffer B .lane 8 The purification of ModA by size-elution chromatography.

Figure 2 Native-PAGE to provide the molybdate lane 1, the modA(acetic buffer PH 5.0); lane 2, the modA incubate with 20mM molybdate.

Sequence and Features