The goldfish hAT-family transposon Tgf2 is capable of autonomous excision in zebrafish embryos

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• 作者：Luo-Dan Cheng ; Xia-Yun Jiang ; Yu-Mei Tian ; Jie Chen ; Shu-Ming Zou
• 关键词：Tc1 ; transposon of Caenorhabditis elegans ; Tgf2 ; transposon of goldfish ; Tol2 ; transposon of Oryzias latipes ; gfTP ; goldfish transposase ; hAT ; hobo of Drosophila ; Ac of maize and Tam3 of snapdragon ; aa ; amino acid(s) ; bp ; base pair(s) ; kb ; kilobase(s) or 1000  ; bp ; mRNA ; message RNA ; ORF ; open reading frame ; PCR ; polymerase chain reaction ; pCS2 ; plasmid containing CMV and SP6 promoter
• 刊名：Gene
• 出版年：15 February, 2014
• 年：2014
• 卷：536
• 期：1
• 页码：74-78
• 全文大小：509 K

文摘

The goldfish (Carassius auratus) Tgf2 transposon is a vertebrate DNA transposon that belongs to the hAT transposon family. In this study, we constructed plasmids containing either the full-length Tgf2 transposon (pTgf2 plasmid) or a partially-deleted Tgf2 transposon (螖pTgf2 plasmid), and microinjected these plasmids into fertilized zebrafish (Danio rerio) eggs at the one- to two-cell stage. DNA extracted from the embryos was analyzed by PCR to assess transient excision, if any, of the exogenous plasmid and to verify whether Tgf2 is an autonomous transposon. The results showed that excision-specific bands were not detected in embryos injected with the 螖pTgf2 plasmid, while bands of 300-500 bp were detected in embryos injected with pTgf2, which indicated that the full-length Tgf2-containing plasmid could undergo autonomous excision in zebrafish embryos. DNA cloned from 24 embryos injected with pTgf2 was sequenced, and the results suggested that Tgf2 underwent self-excision in zebrafish embryos. Cloning and PCR analysis of DNA extracted from embryos co-injected with 螖pTgf2 and in vitro-transcribed transposase mRNA indicated that partially-deleted-Tgf2-containing 螖pTgf2 plasmid also underwent excision, in the presence of functional transposase mRNA. DNA cloned from 25 embryos co-injected with 螖pTgf2 and transposase mRNA was sequenced, and the results suggested that partially-deleted Tgf2 transposons plasmids were excised. These results demonstrated that excisions of Tgf2 transposons were mediated by the Tgf2 transposase, which in turn confirmed that Tgf2 is an autonomous transposon.