Danks Janine A, Ho Patricia M W, Notini Amanda J, Katsis Frosa, Hoffmann Peter, Kemp Bruce E, Martin T John, Zajac Jeffrey D
St Vincent's Institute of Medical Research, Fitzroy, Victoria, Australia.
J Bone Miner Res. 2003 Jul;18(7):1326-31. doi: 10.1359/jbmr.2003.18.7.1326.
A PTH gene has been isolated from the fish Fugu rubripes. The encoded protein of 80 amino acid has the lowest homology with any of the PTH family members. Fugu PTH(1-34) had 5-fold lower potency than human PTH(1-34) in a mammalian cell system.
Parathyroid hormone (PTH) is the major hypercalcemic hormone in higher vertebrates. Fish lack parathyroid glands, but there have numerous attempts to identify and isolate PTH from fish.
Polymerase chain reaction (PCR) was performed with primers based on preliminary data from the Joint Genome Institute database. PCR amplification was performed on genomic DNA isolated from Fugu rubripes. PCR products were purified and DNA was sequenced. All sequence was confirmed from more than one independently amplified PCR product. Multiple sequence alignments were carried out, and the percentage of identities and similarities were calculated. An unrooted phylogenetic tree, using all the known PTH and PTH-related protein (PTHrP) amino acid sequences, was determined. Synthetic peptides were tested in a biological assay that measured cyclic adenosine 3',5'-monophosphate formation in UMR106.1 cells. Rabbit polyclonal antisera specific for N-terminal human PTHrP and one rabbit polyclonal antiserum specific for N terminus hPTH were used to test the cross-reactivity with fPTH(1-34) in immunoblots.
已从红鳍东方鲀中分离出甲状旁腺激素（PTH）基因。编码的含80个氨基酸的蛋白质与任何PTH家族成员的同源性最低。在哺乳动物细胞系统中，红鳍东方鲀PTH（1 - 34）的效力比人PTH（1 - 34）低5倍。
甲状旁腺激素（PTH）是高等脊椎动物中主要的高钙血症激素。鱼类没有甲状旁腺，但人们多次尝试从鱼类中鉴定和分离PTH。
根据联合基因组研究所数据库的初步数据设计引物，进行聚合酶链反应（PCR）。对从红鳍东方鲀分离的基因组DNA进行PCR扩增。对PCR产物进行纯化并测序。所有序列均从多个独立扩增的PCR产物中得到确认。进行多序列比对，并计算同一性和相似性百分比。使用所有已知的PTH和PTH相关蛋白（PTHrP）氨基酸序列构建无根系统发育树。在一种检测UMR106.1细胞中3',5'-环磷酸腺苷形成的生物学测定中测试合成肽。使用针对人PTHrP N端的兔多克隆抗血清和一种针对人PTH N端的兔多克隆抗血清在免疫印迹中检测与红鳍东方鲀PTH（1 - 34）的交叉反应性。
