使用生殖技术创造转基因兔子的方法建议

V. Pokrovskiy, E. A. Patrakhanov, A. Karagodina, Yulia V. Stepenko, N.E. Kazban, A.V. Turpakova, O. Altukhova, P. Lebedev, A. Deykin
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摘要

在这篇文章中,我们提出了一些实用的建议,这些建议是我们在两年的工作中开发出来的,这些工作是通过人工修改基因组获得几系兔子的。通过进一步显微注射基因构建体获得过时的受精卵,评估存活和通过腹腔镜和剖腹手术方法进行胚胎移植,从而获得转基因兔子的技术。以下方案显示出最大的效率:皮下注射血清促性腺激素(Follimag, MOSAGROGEN,俄罗斯),剂量为4ME/kg,静脉注射人绒毛膜促性腺激素(Chorulon,默克动物保健公司,美国),剂量为60ME/kg。因此,由此产生的操作程序允许操作人员以高转移效率和最大的细胞存活率进行显微注射。在视觉控制下,通过NIKON ECLIPSE TS2R倒置研究显微镜,通过Eppendorf FemtoJet 4i气动微注射器使用液压微操纵器进行显微注射。所描述的结果可能有助于编写从事转基因兔动物生产的实验室使用的标准操作程序。
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METHODOLOGICAL RECOMMENDATIONS FOR THE USE OF REPRODUCTIVE TECHNOLOGIES TO CREATE GENETICALLY MODIFIED RABBITS
In this article, we present some practical recommendations that we developed in the course of two years of work on obtaining several lines of rabbits with an artificially modified genome. The technology of obtaining genetically modified rabbits, based on obtaining dated fertilized eggs with further microinjection of genetic constructs, assessment of survival and embryo transfer by laparoscopic and laparotomic methods. The following regimen demonstrated the greatest efficiency: administration of serum gonadotropin (Follimag, MOSAGROGEN, Russia) at a dose of 4ME/kg subcutaneously and intravenous administration of human chorionic gonadotropin (Chorulon, Merck Animal Health, USA) at a dose of 60ME/kg. Thus, the resulting operating procedure allows the operator to perform microinjections with high transfer efficiency and maximum cell survival. Microinjection is carried out through an Eppendorf FemtoJet 4i pneumatic microinjector using hydraulic micromanipulators under visual control through a NIKON ECLIPSE TS2R inverted research microscope. The described results may be useful in the compilation of standard operating procedures used in laboratories that are engaged in the production of genetically modified rabbit animals.
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THE FIRST RECORD OF THE ANTLION MYRMELEON BORE (TJEDER, 1941) (NEUROPTERA: MYRMELEONTIDAE) IN THE SOUTH OF UDMURTIA NEW PALYNOLOGICAL DATA FROM UPPER JURASSIC DEPOSITS OF THE WESTERN CAUCASUS DYNAMICS OF THE WARM SEASON EXTREME TEMPERATURE EVENTS IN IZHEVSK IN THE PERIOD OF 1961-2020 ALL-RUSSIAN CONFERENCE IN RYAZAN METHODOLOGICAL RECOMMENDATIONS FOR THE USE OF REPRODUCTIVE TECHNOLOGIES TO CREATE GENETICALLY MODIFIED RABBITS
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