Kemter E, Philipp U, Klose R, Kuiper H, Boelhauve M, Distl O, Wolf E, Leeb T
Institute for Molecular Animal Breeding and Biotechnology, Gene Center, Ludwig Maximilian University, Munich, Germany.
Cytogenet Genome Res. 2005;111(1):74-8. doi: 10.1159/000085673.
We have cloned the complete coding region of the porcine TNFSF10 gene. The porcine TNFSF10 cDNA has an ORF of 870 nucleotides and shares 85% identity with human TNFSF10, and 75% and 72% identity with rat and mouse Tnfsf10 coding sequences, respectively. The deduced porcine TNFSF10 protein consists of 289 amino acids with the calculated molecular mass of 33.5 kDa and a predicted pI of 8.15. The amino acid sequence similarities correspond to 86, 72 and 70% when compared with human, rat and mouse sequences, respectively. Northern blot analysis detected TNFSF10-specific transcripts (approximately 1.7 kb) in various organs of a 10-week-old pig, suggesting ubiquitous expression. Real-time RT-PCR studies of various organs from fetal (days 73 and 98) and postnatal stages (two weeks, eight months) demonstrated developmental and tissue-specific regulation of TNFSF10 mRNA abundance. The chromosomal location of the porcine TNFSF10 gene was determined by FISH of a specific BAC clone to metaphase chromosomes. This TNFSF10 BAC clone has been assigned to SSC13q34-->q36. Additionally, the localization of the TNFSF10 gene was verified by RH mapping on the porcine IMpRH panel.
我们克隆了猪TNFSF10基因的完整编码区。猪TNFSF10 cDNA有一个870个核苷酸的开放阅读框，与人类TNFSF10的同一性为85%，与大鼠和小鼠Tnfsf10编码序列的同一性分别为75%和72%。推导的猪TNFSF10蛋白由289个氨基酸组成，计算分子量为33.5 kDa，预测的等电点为8.15。与人类、大鼠和小鼠序列相比，氨基酸序列相似性分别为86%、72%和70%。Northern印迹分析在10周龄猪的各种器官中检测到TNFSF10特异性转录本（约1.7 kb），表明其广泛表达。对胎儿期（73天和98天）和出生后阶段（两周、八个月）的各种器官进行实时RT-PCR研究，证明了TNFSF10 mRNA丰度的发育和组织特异性调控。通过将特定BAC克隆与中期染色体进行荧光原位杂交（FISH）确定了猪TNFSF10基因的染色体定位。这个TNFSF10 BAC克隆已被定位到SSC13q34→q36。此外，通过猪IMpRH板上的辐射杂种（RH）图谱验证了TNFSF10基因的定位。
