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Author:
Date:
2015-12-05
[Abstract] Chlamydomonas reinhardtii is a model organism for chloroplast studies. Besides other convenient features, the feasibility of chloroplast genome transformation distinguishes this unicellular alga as ideal for the manipulation of chloroplastic gene expression aiming biotechnological goals, such as improved biofuel and biomass production. Ribulose 1, 5-bisphosphate carboxylase/oxygenase (EC 4.1.1.39, Rubisco) is the photosynthetic carbon-fixing enzyme which is considered crucial for biomass accumulation in algal cultures. Purification of wild type and site-directed mutants of Rubisco in C. reinhardtii is usually performed to study its catalytic properties and assess the carbon-fixing potential of the strains. In this protocol Rubisco is extracted through sonication of cell ...
[摘要] 衣藻衣原体是叶绿体研究的模式生物体。 除了其他方便的特点，叶绿体基因组转化的可行性区分这种单细胞藻理想的操纵叶绿体基因表达瞄准生物技术目标，如改善生物燃料和生物质生产。 核酮糖1,5-二磷酸羧化酶/加氧酶（EC 4.1.1.39，Rubisco）是光合作用碳固定酶，其被认为对藻类培养物中的生物量累积是至关重要的。 Rubisco的野生型和定点突变体的纯化。 通常进行研究其催化性质并评估菌株的碳固定潜力。 在该方案中，通过细胞沉淀的超声处理提取Rubisco，并通过硫酸铵沉淀，蔗糖梯度离心（Goldwaithe和Bogorad，1975）和阴离子交换层析纯化。
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