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Protein molecular weight standard preparation method

A molecular weight standard and protein technology, applied in the field of biochemistry, can solve problems such as high cost and disadvantage, and achieve the effects of low equipment requirements, uniform composition and low cost

Inactive Publication Date: 2015-08-05
HUAQIAO UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Such high costs are detrimental to both producers and users

Method used

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Embodiment 1

[0031] A kind of preparation method of protein molecular weight standard, with elastin-like polypeptide ELP[KV 8 F-20n] as a protein molecular weight standard, where n is an integer or decimal, and 1≤n≤50, where ELP[KV 8 The amino acid sequence of F-20] is shown in SEQ ID 1, and its nucleotide sequence is shown in SEQ ID 2, ELP[KV 8 F-20n] is the above ELP [KV 8 F-20] n-times tandem sequences, that is to say ELP[KV 8 F-20] molecular weight is 9kD, protein molecular weight standard: ELP [KV 8 F-20], ELP[KV 8 F-40], ELP [KV 8 F-80], ELP[KV 8 F-120], ELP [KV 8 F-140], increasing according to the interval gradient of 10kDa or 20kDa.

[0032] The method comprises the steps of:

[0033] (1) Using the recursive directional ligation method (recursive directional ligation, RDL, Meyer D E, Chilkoti A. Genetically encoded synthesis of protein-based polymers with precisely specified molecular weight and sequence by recursive directional ligation: examples from the elastin-like pol...

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PUM

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Abstract

The present invention discloses a protein molecular weight standard preparation method, wherein an elastin-like polypeptide ELP[KV8F-20n] is adopted as a protein molecular weight standard, n is an integer or decimal value and is more than or equal to 1 and is less than or equal to 50, the ELP[KV8F-20] sequence is represented by SEQ ID 1, and the ELP[KV8F-20n] is the tandem sequence formed through tandem connection of the ELP[KV8F-20] n times. According to the present invention, the unique temperature sensitivity of the elastin-like polypeptide is utilized and the high-purity protein can be obtained through the simple centrifugation so as to prepare the protein molecular weight standard; and the method has characteristics of simple operation and low equipment requirement, and is easily subjected to to industrial scale production.

Description

technical field [0001] The invention belongs to the technical field of biochemistry, and in particular relates to a preparation method of a protein molecular weight standard. Background technique [0002] Protein molecular weight standards are necessary biochemical reagents for protein electrophoresis. The usual protein molecular weight standard is a mixture of natural proteins that have been denatured by heating, and the components are not uniform. protein, actin, glyceraldehyde-3-phosphate dehydrogenase, myosin. The preparation of protein molecular weight standards is first of all to obtain high-purity proteins. The procedure for separating and purifying proteins is usually divided into three steps: sample pretreatment, crude fractional purification, and fine fractional purification. The traditional purification procedures mentioned above are relatively complex, time-consuming, require high equipment, and consume a lot of materials, which directly lead to the high cost o...

Claims

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Application Information

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IPC IPC(8): C12N15/70C12N15/11G01N27/447
Inventor 张光亚林源清
Owner HUAQIAO UNIVERSITY
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