Zhao Yaofeng, Pan-Hammarström Qiang, Kacskovics Imre, Hammarström Lennart
Center for Biotechnology, Department of Biosciences at Novum, Karolinska Institutet, Huddinge, Sweden.
J Immunol. 2003 Aug 1;171(3):1312-8. doi: 10.4049/jimmunol.171.3.1312.
The pig delta gene is located approximately 3.4 kb downstream of the second transmembrane exon of the micro gene and shows a similar genomic structure to its counterpart in cow with three exons encoding the CH1, CH2, and CH3 domains. The porcine genomic deltaCH1 exon has been replaced by a recent duplication of the micro CH1 and its flanking sequences, a genetic event that also led to the formation of a short switch delta region, immediately upstream of the delta gene. The deltaCH1 exhibits a 98.7% similarity (314 of 318 bp) to the micro CH1 at the DNA level, whereas the homologies between the deltaCH2 and micro CH3, and the deltaCH3 and micro CH4 are only 33.3 and 35.8%, respectively. Either of the two CH1 exons ( micro and delta) could be observed in the expressed porcine IgD H chain cDNA sequences VDJ- micro CH1-H-deltaCH2-deltaCH3 or VDJ-deltaCH1-H-deltaCH2-deltaCH3, showing a pattern that has not been observed previously in vertebrates. In addition, transfection of a human B cell line, using artificial constructs resembling the porcine C micro -Cdelta locus, also generated both VDJ- micro CH1-deltaCH1-H1-deltaCH2 and VDJ -deltaCH1-H1-deltaCH2 transcripts. An examination of the pig delta genomic sequence shows a putative, second hinge region-encoding exon. Due to the lack of a normal branchpoint sequence for RNA splicing, this exon is not present in the normal pig delta cDNA. However, the exon could be spliced into most of the expressed transcripts in vitro in cell transfection experiments after introduction of a single T nucleotide to restore the branchpoint sequence upstream of the putative H2 exon.
猪δ基因位于微基因第二个跨膜外显子下游约3.4 kb处,其基因组结构与其在牛中的对应物相似,有三个外显子编码CH1、CH2和CH3结构域。猪基因组δCH1外显子已被微基因CH1及其侧翼序列的近期重复所取代,这一遗传事件还导致在δ基因上游紧邻处形成了一个短的转换δ区域。δCH1在DNA水平上与微基因CH1的相似度为98.7%(318 bp中的314 bp),而δCH2与微基因CH3以及δCH3与微基因CH4之间的同源性分别仅为33.3%和35.8%。在表达的猪IgD重链cDNA序列VDJ - 微基因CH1 - H - δCH2 - δCH3或VDJ - δCH1 - H - δCH2 - δCH3中,可以观察到两个CH1外显子(微基因和δ基因)中的任何一个,呈现出一种以前在脊椎动物中未观察到的模式。此外,使用类似于猪C微基因 - Cδ基因座的人工构建体转染人B细胞系,也产生了VDJ - 微基因CH1 - δCH1 - H1 - δCH2和VDJ - δCH1 - H1 - δCH2转录本。对猪δ基因基因组序列的检查显示有一个推测的、编码第二个铰链区的外显子。由于缺乏正常的RNA剪接分支点序列,这个外显子在正常猪δ基因cDNA中不存在。然而,在细胞转染实验中,在引入单个T核苷酸以恢复推测的H2外显子上游的分支点序列后,该外显子可以被剪接到大多数表达的转录本中。