Study On Cloning And Expressing Level Of Flt-1 Gene Of Reproductive Organ In Pregnant Sheep

Vascular epidermal growth factor (VEGF) is a important factor for the growth anddifferentiation of vascular endothelial cells. It plays the important roles in the course of femalemammalian reproduction, including early embryo development; embryo implantation and uterusdegeneration, pregnant maintenance, parturition, and so on. It has the biological activity while itcombined with vascular endothelial growth factor receptor(VEGFR).Studies on theepression and the location of VEGFR was demonstrated in birds porcine,hunman beingand so on, But has not been in Pregnant Sheep. This reports about the study on the femalereproductive organ in the two differren-productivity Ovis aries. Study on the expressing level ofvascular endothelial growth factor receptor—Flt-1 gene of reproductive organ in pregnant sheep.It provided a method for further studying on embryo transplantation, embryonal ectogenesis,treatment of disturbance of genital system of female mammalian.This reports about the study on the female reproductive organ in the Northeast fine wool eweand Small Tail Han adult ewe—uterus, oviduct and ovary. Through Flt-1 gene cDNA libraryconstruction,T-A cloning technique, the products of RT-PCR were ligated with Vector pMD 19-Tand transform E.coli JM109. By the determination of sequence, the recombinent plasmidspMD19-T-VEGFR1 was constructed. Study on the expression of Flt-1 gene of uterus, oviduct andovary in the Northeast fine wool Sheep and Small Tail Han Sheep:β-Actin is dealt with theendogenous internal standard, the RT-PCR technique is used to study the richness of Flt-1 mRNAexpression in different pregnant stages of uteruvs, oviduct and ovary in the Northeast fine woolSheep and Small Tail Han Sheep.The results showed that: (1) The segmental gene of Flt-1 in the uterine horn was by thereverse transcription-polymerase chain reaction (RT-PCR). The length of target gene was 82bp.The results of nucleotide acid sequence analysis showed that the experimental Flt-1 gene has toachieve to 98.9% similarity with the reported Flt-1 genes of the NCBI.(2) In the same pregnant stage, Flt-1 gene had identical expressive tendency in the femalereproductive organ of the adult ewe in the Northeast fine wool Sheep and Small Tail Han Sheep.Flt-1 mRNA was all intensity expression in the endometria of each time. It was thus clear that therewas slight expression in the oviduct and follicle.The expressive intensity strength of Flt-1 mRNA:uterus＞oviduct＞follicle.And the expression of mRNA of Flt-1 was significantly higher in all hystera groups than that of other group (p＜0.05).(3) Flt-1mRNA was exprssed strongly in the uterine horn of the different pregnant stage,and early trimester of pregnancy＜mid trimester of pregnancy＜late trimester of pregnancy.Theexpression of Flt-1 mRNA had gradually fortified tendency. And the expression of mRNA of Flt-1was significantly higher in late trimester of pregnant groups than that of other group (p＜0.05).(4) In the same pregnancy stage, The expression of Flt-1mRNA in the endometria frompregnant Small Tail Han ewe was stronger than pregnant Northeast fine wool ewe. Caption SmallTail Han ewe is hypsi- productivity Ovis aries,and Northeast Fuzz ewe is olig- productivity Ovisaries. The expression of mRNA of Flt-1 did not differ significantly (p＞0.05).