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Prokaryotic Expression and Analysis of McGS1 Gene of Bitter Gourd(PDF)

《北方园艺》[ISSN:1001-0009/CN:23-1247/S]

Issue:
2015年22
Page:
110-113
Research Field:
Publishing date:

Info

Title:
Prokaryotic Expression and Analysis of McGS1 Gene of Bitter Gourd
Author(s):
SHEN Longbin; NIU Yu; LIU Ziji; LIU Zhaohua; YANG Yan
（Tropical Crops Genetic Resources Institute，Chinese Academy of Tropical Agricultural Sciences/Key Laboratory of Crop Gene Resources and Germplasm Enhancement in Southern China，Ministry of Agriculture，Danzhou，Hainan 571737）
Keywords:
bitter gourd (Momordica charantia L.); glutamine synthetase; prokaryotic expression
PACS:
-
DOI:
10.11937/bfyy.201522029
Abstract:
In this study，prokaryotic expression vector expressing bitter gourd McGS1 gene was constructed and transformed into E.coli BL21(DE3) for its expression.A pair of primers was designed according to McGS1 gene sequence.Total RNA of the ‘Re Yan No.3’ bitter gourd was extracted，and then McGS1 gene was amplified by RT-PCR.The recombinant expression vector was identified using enzyme digestion and PCR method.The expression of McGS1 gene was optimized by different concentration of IPTG and induction time.The expression product of McGS1 gene was analyzed by SDS-PAGE.The results showed that the optimal expression condition of McGS1 protein was 37℃，0.2 μmol/L IPTG induction for four hours，and the molecular weight of protein product of McGS1 gene was about 35 kDa.

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Last Update: 2016-01-08