一种新的大鼠红细胞生成素受体基因的剪切形式  

A novel alternative splicing mode of the erythropoietin receptor gene in rats

作　　者：吕秋军[1] 叶棋浓[2] 温利青 高月[1] 孙哲[1] 郭绍明[1] 

机构地区：[1]军事医学科学院放射医学研究所药理毒理室,北京100850 [2]北京生物工程研究所分子遗传研究中心

出　　处：《中华血液学杂志》1999年第11期580-582,共3页Chinese Journal of Hematology

摘　　要：目的　检测大鼠骨髓细胞中新的红细胞生成素受体（ＥｐｏＲ） 基因的可变剪切形式。方法　对正常Ｗｉｓｔａｒ 大鼠骨髓单个核细胞的总ＲＮＡ 进行逆转录聚合酶链反应（ＲＴＰＣＲ） ，并测定扩增片段的序列。结果　用一对引物（５′引物为６３７ ～６５７ｂｐ，３′引物为８８２～９０３ ｂｐ） 可扩增出两条片段，一条长约为２６７ ｂｐ，另一条为较长的片段。其中２６７ ｂｐ 的片段为正常的ＥｐｏＲ扩增片段。测序结果显示，较长的扩增片段长度为３４６ ｂｐ，同样为大鼠ＥｐｏＲ 核苷酸序列，但在７３６ 和７３７ 碱基之间插入了长度为７９ ｂｐ的片段。经序列检索发现，７９ｂｐ 的插入序列为滞留的第Ⅴ内含子，但在插入序列的第１６ 位碱基由Ｃ突变为Ｔ，第４９ 位和第５０ 位碱基之间插入了Ｇ 碱基。上述移码突变导致基因的阅读框架发生变化，在第７ 外显子出现终止密码子。结论　在正常的Ｗｉｓｔａｒ 大鼠中发现了新的、胞外近膜区、跨膜区和截短的胞内区编码序列发生变化的ＥｐｏＲｍＲＮＡ 可变剪切形式。这一可变剪切形式的生物学意义有待于进一步研究。Objective To detect new alternative splicing mode of erythropoietin receptor (EpoR)gene in normal rats. Methods Total RNAs were isolated from bone marrow mononuclear cells of normal Wistar rats. Reverse transcriptase polymerase chain reaction (RT PCR) of the total RNAs was done and the amplified products were sequenced. Results After RT PCR with a pair of primers (sense: 637/657 bp, and antisence: 903/882 bp), two bands were detected, the expected band (267?bp) and a longer one (346?bp) which had a 79?bp insert between 736th and 737th bases from the first ATG of the rat EpoR nucleotide. Sequence analysis showed that the 79?bp insert was the retaining intron 5 with a C to T point mutation at 16th base and an insertion of G base between 49th and 50th base. The insertion of G base caused a shift in reading frame and resulted EpoR with a truncated intracellular domain. Conclusion A new alternative splicing of EpoR mRNA encoding truncated intracellular domain was identified in normal Wistar rats. Its implication in rat erythropoiesis remains to be determined.

关 键 词：受体 红细胞生成素 基因 聚合酶链反应 

分 类 号：R331.14[医药卫生—人体生理学]