Extraction method of ginkgo leaf chloroplast protein

Abstract

The invention belongs to the field of biological chemistry and specifically relates to extraction method of chloroplast protein of xylophyta ginkgo leaf. The method comprises the following steps of: firstly, preparing coarse chloroplast particles from the ginkgo leaves, performing gradient centrifugation through Percoll, so as to obtain complete chloroplast by a specific centrifugal force; then, dissolving protein by using a protein extraction buffer solution, adding the same volume of Tris-balanced phenol for extraction, and precipitating protein by using a ammonium acetate methanol solution. The extraction method is capable of effectively increase extraction efficiency and purity of chloroplast protein. The ginkgo leaf chloroplast protein obtained by the method can be used for proteomics analysis of ginkgo leaf and other xylophyta materials rich in interfering substances, so that solubility and purity of protein are improved; a 2-DE map of the protein has a clear background and many protein spots are distinguishable.

Description

technical field [0001] The invention belongs to the field of biochemistry, and in particular relates to a method for extracting chloroplast protein from leaves of woody plant ginkgo biloba. Background technique [0002] Ginkgo biloba L. is a deciduous tree of the genus Ginkgo in the Ginkgo family. It is known as the "living fossil" in the plant kingdom and is the oldest gymnosperm among the existing seed plants. China is the hometown of ginkgo, and the existing ginkgo resources account for more than 70% of the world's ginkgo resources. Ginkgo biloba is an excellent economic tree species integrating food, medicine, health care, cosmetics and materials [1] In recent years, ginkgo has also been widely used as an important greening tree species all over the country. Because Ginkgo biloba is a precious relict plant, it has many original characteristics. It is one of the plant varieties that scientists focus on and has been valued and studied by different scientific researchers....

AI Technical Summary

Problems solved by technology

It can be seen that the preparation of chloroplast protein samples of ginkgo leaves is more difficult

TCA-acetone precipitation is currently the most common method for protein extraction. Although the operation steps are relatively simple, the yield of protein crude extract is relatively high. [5] , but the protein powder obtained by this method has poor resolubility when extracting Ginkgo chloroplast protein, and there are more impurities in the sample, and the background of the 2-DE map is blurred and there will be obvious stripes [6]

However, the current research on plant chloroplast protein extraction methods mainly focuses on herbaceous plants such as rice and Arabidopsis. [7] , while Ginkgo biloba is a precious relict woody plant, the efficiency and quality of its chloroplast protein extraction are not enough to meet the needs of its proteomics research

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