GENES ANCESTRAL TO THE THYROID/STEROID RECEPTOR FAMILY

为了更好地理解生物多样性的功能和进化 类固醇/甲状腺(S/T)激素受体基因大家族 一直在调查它们在低等生物中的存在。我们是在 在酵母菌中找不到它们，但在蛇床子虫和 线虫。 对秀丽隐杆线虫的线虫研究工作已在 与迈克·克劳斯博士的合作。我们之前克隆了三个cDNA 从线虫、chr3、cnr8和cnr14中克隆了 并对这些DNA的基因进行了测序。CHR 3有8个外显子和7个外显子 内含子。我们还有3,500个碱基的序列 转录起始点。利用这一点，我们构建了一个带有 Lac Z是一名记者。通过以下方式获得转基因蠕虫是可能的 将这个结构注入卵巢。染色显示有 在胚胎发育早期的皮下细胞中是1.5倍 阶段，并延伸到成虫。这一点得到了以下人士的证实 抗lin26抗体与chr3的共定位。众所周知，LIN26是 仅在皮下细胞中表达。Chr3的杂交 带有skin1突变的转基因蠕虫进一步 确认皮下细胞定位，因为这些蠕虫显示chr3 在这种突变引起的异常皮下细胞中的定位。 我们目前正在尝试用一种抗体进行定位 Chr3蛋白。我们还试图获得转基因蠕虫 Cnr14基因。 在我们工作的另一部分中，我们利用了几种形式的线虫。 第一种是三角草属(Tripealia Cystophora)。在这里，我们隔离了 对S/T基因进行了部分测序，该基因以两种异构体和 属于RXR家族。我们还检查了另一种蛇怪， 可在培养条件下生长的金鱼藻。在这个有机体中，我们 已经分离并部分测序了三个不同的S/Y基因。 这些代表了孤儿受体RAR-RXR的同源物和 蜕皮激素受体。后者是有意义的，因为 从息肉到水母的发育(花柱状)是一种 蜕变。我们已经成功地采集了息肉并分解了 它们变成单个细胞，然后重新聚集，看着它们形成息肉 再来一次。我们目前正试图在这一领域制造转基因的奥雷利亚 S/T受体在此过程中的方式和作用的研究 跳跃的声音。

In an attempt to understand better both the functions and evolution of the large family of steroid/thyroid (S/T) hormone receptor genes we have been investigating their presence in lower organisms. We were unable to find any in yeast bu have found them in cnidaria and nematodes. The nematode work with Caenorhabditis elegans has been carried on in collaboration with Dr. Mike Krause. We previously cloned three cDNAs from C. Elegans, chr3, cnr8 and cnr 14. Subsequently, we have cloned and sequenced the genes for these cDNAs. Chr 3 has eight exons and 7 introns. We also have the sequence of 3,500 bases upstream of the transcription start site. Using this we constructed a plasmid with Lac Z as a reporter. It was possible to obtain transgenic worms by injecting this construct into the ovary. Staining showed expression in hypodermal cells beginning early in embryogenesis at the 1.5 fold stage and extending into the adult. This was confirmed by colocalization of chr3 with antibodies to lin26. Lin26 is known to be expressed exclusively in hypodermal cells. Crossing of the chr3 transgenic worms with worms bearing the skin1 mutation further confirmed hypodermal cell localization since these worms showed chr3 localization in the aberrant hypodermal cells caused by this mutation. We are currently attempting to obtain localization with an antibody to the chr3 protein. We are also attempting to obtain worms transgenic for the cnr14 gene. In the other part of our work we utilize several forms of cnidaria. The first is Tripedalia cystophora. Here we have isolated and partially sequenced a single S/T gene which exists in two isoforms and belongs to the RXR family. We also examined another cnidarian, Aurelia aurita which can be grown in culture. In this organism we have isolated and partially sequenced three different S/Y cDNAs. These represent homologues of RAR-RXR, an orphan receptor and the ecdysone receptor. The latter is of interest because the process of development from polyp to medusa (strobilation) is a type of metamorphosis. We have successfully taken polyps and disaggregated them into single cells, reaggregated then and seen them form polyps again. We are currently attempting to make transgenic aurelia in this manner and investigating the role of the S/T receptors in the process of strobilation.