早期胚胎血管显微注射慢病毒载体制备转基因禽类的研究

doi:10.11843/j.issn.0366-6964.2015.12.009

Abstract

Abstract:

This study was aimed to identify the availability of early embryo blood vessels microinjection of lentiviral vector as a new，effective way in making transgenic birds，here，we combined some of the transgenic technology such as primordial germ cells(PGCs)，microinjection and lentiviral vector together according to the characteristic of bird development.The number of PGCs in the bloodstream was maximal when the chicken embryoes developed to Hamburger-Hamilton stage 13-15(HH13-15)，1 μL lentiviral vector，containing an enhanced-green fluorescent protein(eGFP)，was microinjected into the blood vessels of quail embryos at stage HH13-15 with a titer of 1×10⁹TU•mL^-1.A total of 80 embryos were injected and 48 quails(60%) were successfully hatched.In newly hatched quails，eGFP expression was shown as the presence of green fluorescence in the beak，feather，eye，brain，blood vessels，heart，liver，spleen，lung，kidney，glandular stomach，mesenterium，small intestine，large intestine，oviduct，muscle and claws，especially in gonads.In 5 out of 21 mature G0 male quails，the semen was eGFP-positive(5/21，23.8%)，as detected by polymerase chain reaction(PCR).Southern blot and genetic analyses revealed that in the 46 G1 offspring produced by G0 quail，6 were transgenic(6/46，13.0%) according to the PCR and Southern blot results.In conclusion，transgenic germ line chimeras was successfully generated by injection of lentiviral vector into embryonic blood vessel at stage HH 13-15.It indicated that infection of PGCs with lentiviral vector via direct injection into blood vessels has the potential to provide a more convenient and efficient way to produce transgenic birds.

CLC Number:

S814.8

ZHANG Zi-fu，ZHAO Yu，LIU Jin-ni，LI Xun，PENG Xin-liang，ZHANG Ping，HU Jing，HE Dong-yang，CHANG Jiao-jiao，GU Meng-di，JIN Nan-nan，WANG Ya-fang. Producing Transgenic Avian by Microinjection of Lentiviral Vector into the Early Embryo Blood Vessels[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(12): 2185-2191.

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Producing Transgenic Avian by Microinjection of Lentiviral Vector into the Early Embryo Blood Vessels

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