首页> 中文期刊>南方医科大学学报 >一种新的人血管内皮生长因子基因的剪切体

一种新的人血管内皮生长因子基因的剪切体

     

摘要

Objective To characterize a new alternative splicing isoform of human vascular endothelial growth factor (VEGF)gene.Methods The total RNA was extracted from the lung tissue of a legally aborted 4-month-old fetus and amplified by RT-PCR.The amplified product was cloned into the plasmid pMD18-T and plasmid pcDNA3.1- for sequence analysis.Results Electrophoresis of the RT-PCR products displayed one short band for VEGF121 (487 bp) and a long band.The latter was dharacterized to contain two fragments:one was normal VEGF165 (619 bp),and the other (639 bp) had an identical nucleotide sequence to VEGF165 with a 20 bp fragment inserted between exons 3 and 4.Sequence analysis showed that this 20-bp nudeotide was inserted from the 3' end of the third intron containing a splicing signal thus causing shift mutation in the reading frame of VEGF gene and early appearance of the stop codon UAG in the middle of exon 4.Conclusion A new altemative splicing isoform of VEGF probably exists in the lung tissue of a legally aborted human fetus,and its biological significance remains to be further investigated.%目的 探索人胚胎肺组织中是否存在一种新的血管内皮生长因子(VEGF)基因可变剪切形式.方法 对1例4月龄引产胎儿肺组织的总RNA进行逆转录聚合酶链反应、克隆并测定扩增片段的序列.结果 用一对引物(引物5’端-21~7 bp,3’端554~576 bp)可扩增出3条片段,一条为长度正常的VEGF165(619 bp)全长序列,另一条为长度正常的VEGF121(487bp)全长序列,第三条为新的VEGF mRNA异常剪切片段.测序检测显示,此扩增片段长度为639 bp,同样为VEGF165全长核苷酸序列,但在VEGF165的第3和第4外显子之间插入了长度为20bp的碱基片段.序列分析显示此插入序列为滞留的第Ⅲ内含子终末序列,含内含子剪切信号.上述移码突变导致VEGF基因阅读框架发生改变,在第4外显子中游出现终止密码,故仅含-VEGF165外显子l~3及4部分序列,在蛋白水平上可能编码含97个氨基酸的新肽链.结论 在一引产胎儿的肺组织中发现了新的VEGF mRNA可变剪切形式,其生物学意义有待进一步研究.

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