piggyBac转座子在绒山羊基因组中的整合位点及其特征分析

doi:10.3864/j.issn.0578-1752.2012.05.017

Abstract

Abstract: 【Objective】 The objective of the experiment was to construct piggyBac (PB) transposons vectors and integration site in Cashmere goat genome of PB was also studied. 【Method】 Donor plasmid of PB-CMV-EGFP-Neo and helper-dependent plasmid of pcDNA-Trans were constructed and transferred into Cashmere goat fetal fibroblasts by lipofectamine 2000. Cell clones stably transfected were obtained after screening by G418. Transgenic cell genome was obtained and the integration site of PB transposons were detected by r-PCR.【Result】Transposition system was constructed and the integration of foreign gene in Cashmere goat fibroblasts was successfully mediated. Transgenic cell line was also obtained. There were 21 integration sites of PB transposon in Cashmere goat genome after r-PCR detection. Analysis of the composition of the five bases, which was close to the side of the PB integration sites TTAA, showed that PB 3′ tended to insert into region rich in AT (58.35%) and the PB 5′ tended to insert into region rich in GC (57.8%). 【Conclusion】 The transposition of foreign PB transposons in Cashmere goats was highly efficient. The integration site information which is acquired from this research will provide theoretical references for Cashmere goats study by PB transposons.

Key words: piggyBac, Cashmere goat, genome, integration sites

BAI Ding-Ping, FANG Kun, YANG Ming-Ming, QU Lei, CHEN Yu-Lin. Integration Sites and Their Characteristic Analysis of piggyBac Transposon in Cashmere Goat Genome[J].Scientia Agricultura Sinica, 2012, 45(5): 958-965.

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Integration Sites and Their Characteristic Analysis of piggyBac Transposon in Cashmere Goat Genome

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