The invention relates to high-level production of pHBA in green plants using a unique expression cassette. The latter comprises a chorismate pyruvate lyase (CPL) coding sequence operably linked to a suitable promoter capable of driving protein expression in higher plants. Additionally, the CPL cassette comprises a sequence encoding a chloroplast transit peptide, its natural cleavage site, and a small portion of the transit peptide donor protein fused to the N-terminus of CPL. The chloroplast targeting sequence targets the foreign protein to the chloroplast compartment and aids in its uptake into the organelle. The cleavage site is unique to the transit peptide, and cleavage of the chimeric protein encoded by the cassette at this site releases a novel polypeptide that has full enzyme activity, comprising the mature CPL enzyme and a small portion of the transit peptide donor.
本发明涉及使用独特的表达盒在绿色植物中高
水平生产pHBA。该表达盒包括一个编码脱氧
香豆酸-
丙酮酸裂解酶(
CPL)的序列,与能够在高等植物中驱动蛋白质表达的适当启动子可操作地连接。此外,
CPL表达盒包括一个编码叶绿体转运肽的序列,其天然裂解位点和一小部分转运肽供体蛋白与
CPL的N-端融合。叶绿体定位序列将外源蛋白定位到叶绿体区域并有助于其进入细胞器。裂解位点是转运肽唯一的,该盒编码的嵌合蛋白在该位点的裂解释放出一个全酶活性的新型
多肽,包括成熟的
CPL酶和一小部分的转运肽供体。