胸腺肽基因多元植物表达载体的构建

Polyphyletic Expressing Vector Construction of Thymosin Gene and Transformation to Carrot Callous

Gus基因是最为常用的报告基因之一,为了便于对胸腺肽目的基因进行检测,在该研究中,胸腺肽目的基因、PRIB分泌基因和GUS报告基因DNA片段用DNA纯化回收试剂盒回收,然后用T4 DNA连接酶连接,构建成几个基因相融合的多元植物表达载体,并通过酶切进行鉴定。用基因枪转化法对胡萝卜愈伤组织进行了遗传转化,转化的胡萝卜愈伤组织经Southern杂交检测和X-Gluc染色,结果表明,多元表达载体成功地被导入胡萝卜愈伤组织。

Gus gene is one of the most popular used reporter genes.β-glucuronidase gene,PR1b secreting gene and thymosin gene segments were reclaimed and constructed into polyphyletic expressing vectors and the thymosin gene could be detected easily.The polyphyletic expressing vectors were detected by endonuclease digestion.Carrot callus was transformed using the polyphyletic vectors by bombardment The transformed callus was detected to be blue by X-Gluc tinction,and to be positive by Southern blotting.The results indicated that the polyphyletic expressing vector was transferred into genome of carrot callous.