An efficient extraction method of chloroplast proteins from ginkgo plants compatible for two-dimensional electrophoresis

doi:10.3969/j.issn.1000-2006.201604047

JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2017, Vol. 41 ›› Issue (03): 51-56.doi: 10.3969/j.issn.1000-2006.201604047

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An efficient extraction method of chloroplast proteins from ginkgo plants compatible for two-dimensional electrophoresis

SHI Dawei^1,2, HE Mei^2,3, LIANG Feifei², XU Ming², XIE Yinfeng^1,2

1. Co-Innovation Center for the Sustainable Forestry in Southern China, Nanjing 210037, China;
2. College of Biology and the Environment, Nanjing Forestry University, Nanjing 210037, China;
3. Jiangxi Academy of Forestry, Nanchang 310032, China

Online:2017-06-18 Published:2017-06-18

Abstract

Abstract: 【Objective】To explore the mechanism of the response of photosynthesis in ginkgo plants at protein level to environmental conditions, an extraction method for chloroplast proteins was investigated. 【Method】An experimental system was developed that was compatible with two-dimensional electrophoresis(2-DE)of ginkgo chloroplast proteins. Tris-phenol method was used for the extraction of ginkgo chloroplast proteins, and compared with trichloroacetic(TCA)-acetone precipitation method.【Result】The results indicated that Percoll step gradient centrifugation was suitable for ginkgo chloroplast isolation, and the mean intact rate of purified chloroplasts was approximately 85%. The results of one-dimensional PAGE indicated that protein bands of chloroplasts were more distinct at the low molecular weight region of the gel for the Tris-phenol extracti method compared with the TCA-acetone precipitation method. Moreover, comparison of this method with TCA-acetone precipitation for the extraction of ginkgo chloroplast proteins showed that the method resulted in higher protein yields and produced more protein spots on 2-DE gels with less streak interference. 【Conclusion】The above results revealed that the Tris-phenol extraction method could efficiently extract high-quality chloroplast proteins for 2-DE, and could be applied in proteomic analysis of ginkgo chloroplasts.

CLC Number:

S718.3

SHI Dawei, HE Mei, LIANG Feifei, XU Ming, XIE Yinfeng. An efficient extraction method of chloroplast proteins from ginkgo plants compatible for two-dimensional electrophoresis[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY, 2017, 41(03): 51-56.

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An efficient extraction method of chloroplast proteins from ginkgo plants compatible for two-dimensional electrophoresis

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