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家蚕中 PB 转座子的不可预测重组:潜在风险。

Unpredictable recombination of PB transposon in Silkworm: a potential risk.

机构信息

State Key Laboratory of Silkworm Genome Biology, College of Textile and Garment, Southwest University, No. 2 Tiansheng Street, Beibei, Chongqing, 400715, China.

Chongqing Engineering Research Center of Biomaterial Fiber and Modern Textile, Chongqing, 400715, China.

出版信息

Mol Genet Genomics. 2021 Mar;296(2):271-277. doi: 10.1007/s00438-020-01743-0. Epub 2020 Nov 17.

Abstract

The piggyBac (PB) transposon is the most widely used vector for generating transgenic silkworms. The stability of the PB transposon in the receptor is a serious concern that requires attention because of biosafety concerns. In this study, we found that the transgene silkworm developed loss of reporter gene traits. To further investigate the regularity, we traced the genes and traits of this silkworm. After successful alteration of the silkworm genome with the MASP1 gene (named red-eyed silkworm; RES), silkworm individuals with lost reporter genes were found after long-term transgenerational breeding and were designated as the white-eyed silkworm (WES). PCR amplification indicated that exogenous genes had been lost in the WES. Testing was conducted on the PB transposons, and the left arm (L arm) did not exist; however, the right arm (R arm) was preserved. Amino acid analysis showed that the amino acid content of the WES changed versus the common silkworm and RES. These results indicate that the migration of PB transposons in Bombyx mori does occur and is unpredictable. This is because the silkworm genome contains multiple PB-like sequences that might influence the genetic stability of transgenic lines. When using PB transposons as a transgene vector, it is necessary to fully evaluate and take necessary measures to prevent its re-migration in the recipient organism. Further experiments are needed if we want to clarify the regularity of the retransposition phenomenon and the direct and clear association with similar sequences of transposons.

摘要

转座子 piggyBac (PB) 是用于生成转基因蚕的最广泛使用的载体。由于生物安全问题,受体中转座子 PB 的稳定性是一个需要关注的严重问题。在这项研究中,我们发现转基因蚕失去了报告基因的特性。为了进一步研究其规律,我们追踪了这种蚕的基因和特性。在用 MASP1 基因(命名为红眼蚕;RES)成功改变蚕的基因组后,在长期的转代繁殖中发现了失去报告基因的蚕个体,并将其命名为白眼蚕(WES)。PCR 扩增表明 WES 中外源基因已丢失。对 PB 转座子进行了测试,发现其左臂(L 臂)不存在,而右臂(R 臂)则保留下来。氨基酸分析表明,WES 的氨基酸含量与普通蚕和 RES 相比发生了变化。这些结果表明,PB 转座子在家蚕中的迁移确实会发生,而且是不可预测的。这是因为家蚕基因组中含有多个 PB 样序列,可能会影响转基因系的遗传稳定性。当使用 PB 转座子作为转基因载体时,有必要充分评估并采取必要措施来防止其在受体生物中的重新迁移。如果我们要阐明重转座现象的规律以及与转座子相似序列的直接明确关联,还需要进一步的实验。

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