本发明涉及生物基因工程技术领域，是一种构建等鞭金藻叶绿体同源重组转基因系统的方法。以等鞭金藻叶绿体基因组来源的SEQ ID NO : 1所示序列和SEQ ID NO : 2所示的序列为同源臂，SEQ ID NO : 3所示的序列、SEQ ID NO : 4所示的序列为启动子，以SEQ ID NO : 5所示的序列、SEQ ID NO : 6所示的序列为终止子，以bar基因为筛选标记基因，并通过SEQ ID NO : 7连接多个外源基因构成多顺反子结构从而构建同源重组载体，并利用基因枪法将载体导入等鞭金藻细胞，经除草剂草丁膦筛选获得转基因藻株。采用本发明的等鞭金藻叶绿体同源重组转基因系统, 可实现多个外源基因在叶绿体中稳定表达。
其他摘要
The invention relates to the technical field of biological genetic engineering, in particular to a method for constructing an isochrysis sp. chloroplast homologous recombinant transgenic system. Sequences shown as SEQ ID NO : 1 and SEQ ID NO : 2 derived from an isochrysis sp. chloroplast genome are taken as homologous arms; sequences shown as SEQ ID NO : 3 and SEQ ID NO : 4 are taken as promoters; sequences shown as SEQ ID NO : 5 and SEQ ID NO : 6 are taken as terminators, a bar gene is taken as a screening marker gene, a plurality of exogenous genes are connected through SEQ ID NO : 7 to form a polycistron structure so as to construct a homologous recombinant vector, the vector is introduced into isochrysis sp. cells by using a gene gun method, and a transgenic algal strain is obtained through herbicide glufosinate-ammonium screening. By adopting the isochrysis sp. chloroplast homologous recombinant transgenic system, a plurality of exogenous genes can be stably expressed in chloroplast.
; 王寅初
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