Scientia Agricultura Sinica ›› 2012, Vol. 45 ›› Issue (5): 958-965.doi: 10.3864/j.issn.0578-1752.2012.05.017

• ANIMAL SCIENCE·RESOURCE INSECT • Previous Articles     Next Articles

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

 BAI  Ding-Ping, FANG  Kun, YANG  Ming-Ming, QU  Lei, CHEN  Yu-Lin   

  1. 1.西北农林科技大学动物科技学院，陕西杨凌 712100
    2.新疆农垦科学院畜牧兽医研究所，新疆石河子 832000
    3.榆林学院陕西省陕北绒山羊工程技术研究中心，陕西榆林 719000
  • Received:2011-04-06 Online:2012-03-01 Published:2011-07-19

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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

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