CAJ | 학술논문

以小球藻及莱茵衣藻原生质体为受体细胞，利用电击法将质粒 pCAMBIA1301转入小球藻和莱茵衣藻，摸索电击转化条件并进行分子检测。结果表明：两类藻都对潮霉素敏感，小球藻及莱茵衣藻分别在含25 mg／L和100 mg／L潮霉素的固体培养基上的生长被完全抑制；小球藻和莱茵衣藻原生质体电击转化的最佳电击场强分别为0．8 kV／cm和0．6 kV／cm，最佳脉冲时间均为10 ms；制备原生质体和通过2-脱氧-D-葡萄糖处理可明显提高转化效率；分子检测说明GU S报告基因成功转入两种藻并可稳定遗传。
이소구조급래인의조원생질체위수체세포，이용전격법장질립 pCAMBIA1301전입소구조화래인의조，모색전격전화조건병진행분자검측。결과표명：량류조도대조매소민감，소구조급래인의조분별재함25 mg／L화100 mg／L조매소적고체배양기상적생장피완전억제；소구조화래인의조원생질체전격전화적최가전격장강분별위0．8 kV／cm화0．6 kV／cm，최가맥충시간균위10 ms；제비원생질체화통과2-탈양-D-포도당처리가명현제고전화효솔；분자검측설명GU S보고기인성공전입량충조병가은정유전。
The plasmid pCAMBIA1301 was transformed into The receptor cells Chlorella vulgaris and Chlamydomonas reinhardtii protoplasts by electroporation method.The related parameters was tested and molecular detections were per-formed in transformants.The results showed that:two kinds of algae were sensitive to hygromycin.The growth of Chlo-rella and Chlamydomonas was completely inhibited on solid media containing 25 mg/L and 100 mg/L concentration of hygromycin respectively.The optimal electric field intensity were 0.8 kV/cm and 0.6 kV/cm in Chlorella and Chlamydomonas respectively.The optimal pulse time was 10 ms for both species.Making protoplasts and treating with 2-deoxy-D-glucose make it possible to significantly increase the transformation rates.Molecular identification indicated that GUS gene has been successfully transformed into the two species of algae and the genetic form is stable.