Ashkani Jahanshah, Rees D J G
Biotechnology Department, University of the Western Cape, Robert Sobokwe Road, Bellville, 7535, South Africa.
Agricultural Research Council, Biotechnology Platform, Private Bag X5, Onderstepoort, 0110, South Africa.
Biotechnol Rep (Amst). 2017 Jun 16;15:41-47. doi: 10.1016/j.btre.2017.06.005. eCollection 2017 Sep.
A single locus ( locus) including at least two linked genes (female and male determinants) genetically controls the gametophytic self-incompatibility (GSI) in apple, which has evolved to avoid self-fertilization. There has been extensive work done on the female determinant of self-incompatibility, which has led to the determination of the tertiary structure of -RNase. However, the tertiary structure of male determinant ( locus F-box, SLF/SFB) remains unresolved, which could mainly be due to difficulties associated with its expression in the recombinant expression systems. In addressing this, we have evaluated several (prokaryotic and eukaryotic) and expression systems for their efficiency in the expression of apple SLF2. The most successful expression of SLF2 (1 mg/ml) was achieved in using the synthesized gene in a high salt culture and applying heat shock before induction of culture. We therefore present an approach for the efficient expression of locus F-box recombinant proteins for future functional and structural studies.
一个单一位点（位点），包括至少两个连锁基因（雌性和雄性决定因子），从基因层面控制苹果中的配子体自交不亲和性（GSI），这一特性的进化是为了避免自花受精。关于自交不亲和性的雌性决定因子已经开展了大量研究工作，这使得-RNase的三级结构得以确定。然而，雄性决定因子（位点F-box，SLF/SFB）的三级结构仍未解析，这主要可能是由于其在重组表达系统中表达存在困难。为了解决这个问题，我们评估了几种（原核和真核）表达系统表达苹果SLF2的效率。通过在高盐培养基中使用合成基因并在诱导培养前进行热休克处理，在中实现了SLF2的最成功表达（1毫克/毫升）。因此，我们提出了一种有效表达位点F-box重组蛋白的方法，用于未来的功能和结构研究。
