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稀有鮈鲫(Gobiocypris rarus)是我国特有的一种小型鲤科鱼类,目前已作为一种新型模式生物应用于实验研究。运用普通PCR方法克隆得到一段长6 299 bp的序列,序列分析发现编码区与稀有鮈鲫卵黄蛋白原cDNA序列相似性为100%。再参考6 299 bp序列5’端设计两对特异性引物,进行基因组步移,获得长度为1 285 bp的序列。经过序列拼接后得到7 573 bp的序列,分析发现其中含卵黄蛋白原基因全序列长度为6 595 bp卵黄蛋白原基因上游调控序列长度为978 bp。进一步分析开放性阅读框得知:稀有鮈鲫卵黄蛋白原基因含有26个外显子和25个内含子,编码1 299个氨基酸序列,CDS长3 900 bp。经BLAST比对发现,稀有鲫卵黄蛋白原上游调控序列中220~838 bp区间与朝鲜(Acheilognathus yamatsutae)卵黄蛋白原基因启动子序列2 857~3 469 bp区间有80%相似性。
Gobiocypris rarus, a small carp species endemic to China, has been used as a new model organism in experimental studies. A sequence of 6 299 bp was cloned by ordinary PCR. The sequence analysis showed that the similarity of the cDNA sequence between the coding region and the yolk protozoan of the crucian carp was 100%. Reference to the 6 299 bp sequence 5 ’end of the design of two pairs of specific primers for genome walking to obtain a length of 1 285 bp sequence. After sequencing, a 7 573 bp sequence was obtained. The result showed that the full length of the yolkinogen gene sequence of 6955 bp was 978 bp. Further analysis of the open reading frame revealed that the rare yak carp yolk protozoa contained 26 exons and 25 introns, encoding 1 299 amino acid residues and the CDS length was 3 900 bp. The results of BLAST showed that the sequence of 220-838 bp in the upstream regulatory sequence of yolk primordium was 80% similar to that of the genus Acheilognathus yamatsutae in the range of 2 857-3 469 bp.