发育特异基因及其在RNA水平上的表达

DEVELOPMENT-SPECIFIC GENES AND THEIR EXPRESSION AT THE RNA LEVEL

从λ噬菌体gt11的cDNA表达文库筛选到7个胡萝卜体细胞发育特异的基因克隆,并对它们进行了纯化、分离噬菌体DNA,限制酶消化,琼脂糖凝胶电泳鉴定,以及将cDNA插入片段再克隆到pIBI或p^(UC18)质粒等处理。然后,通过Southern blot分析,证明克隆16,22,50及60等为新的cDNA基因。此外,又通过Northern blot分析测定了这些基因的组织特异性表达及时序表达。结果表明,克隆22是一种受发育调节,与胡萝卜体细胞胚胎形成密切相关的基因。

A total of seven clones of development-specific genes were screened from the cDNA expression library of lambda gtll constructed from carrot somatic embryos. The purification of plaques, isolation of DNA from the phages, digestion of the clone's DNA with certain restriction enzyme, identification of the inserts on agarose gel by electrophoresis, and subcloning of cDNA inserts from lambda gtll. into plasmid pIBI or pUC18 were performed, respectively.Date of cross hybridization by Southern blot analysis indicated that clones 16, 22, 50 and 60 are new cDNA genes. In addition, the new cDNA gene's tissue-specific expression on variety of organs in the carrot plant, such as flower, petiole, leaf and root as well as time-course expression in callus and embryo cells of different culture periods were determined. The results suggest that the corresponding gene for clone 22 is a specific gene regulated by development and closely related to carrot somatic embryogenesis.