Hallmann A, Wodniok S
Department of Cellular and Developmental Biology of Plants, University of Bielefeld, Universitätsstr 25, 33615 Bielefeld, Germany.
Plant Cell Rep. 2006 Jun;25(6):582-91. doi: 10.1007/s00299-006-0121-x. Epub 2006 Feb 3.
Production of transgenic organisms is a well-established, versatile course of action in molecular biology. Genetic engineering often requires heterologous, dominant antibiotic resistance genes that have been used as selectable markers in many species. However, as heterologous 5' and 3' flanking sequences often result in very low expression rates, endogenous flanking sequences, especially promoters, are mostly required and are easily obtained in model organisms, but it is much more complicated and time-consuming to get appropriate sequences from less common organisms. In this paper, we show that aminoglycoside 3'-phosphotransferase gene (aphVIII) based constructs with 3' and 5' untranslated flanking sequences (including promoters) from the multicellular green alga Volvox work in the unicellular green alga Chlamydomonas and flanking sequences from Chlamydomonas work in Volvox, at least if a low expression rate is compensated by an enforced high gene dosage. This strategy might be useful for all investigators that intend to transform species in which genomic sequences are not available, but sequences from related organisms exist.
转基因生物的生产是分子生物学中一种成熟且通用的操作方法。基因工程通常需要异源的显性抗生素抗性基因，这些基因已在许多物种中用作选择标记。然而，由于异源的5'和3'侧翼序列往往导致极低的表达率，因此大多需要内源侧翼序列，尤其是启动子，并且在模式生物中很容易获得，但从不太常见的生物中获取合适的序列则要复杂得多且耗时。在本文中，我们表明，基于氨基糖苷3'-磷酸转移酶基因（aphVIII）构建的载体，其带有来自多细胞绿藻团藻的3'和5'非翻译侧翼序列（包括启动子），能在单细胞绿藻衣藻中发挥作用，而来自衣藻的侧翼序列也能在团藻中发挥作用，至少在通过强制提高基因剂量来补偿低表达率的情况下如此。该策略可能对所有打算转化那些没有基因组序列但存在相关生物序列的物种的研究人员有用。
