| Telomerase is a specialized cellular ribonucleoprotein (RNP) reverse transcriptase (RT), with core enzyme containing the protein subunit and the RNA component. Telomeres lost during DNA replication can be added back by the activity of telomerase, regulated in the mammalian development. The telomere rebuilding may be attributed to the reprogramming of telomerase activity at the different stages in the early embryo development. Therefore, investigating telomere and telomerase reprogramming during early embryo development is of great importance.In this study,Yanbian Yellow cattle as the materials and used the 293-cell as the standard. While,detection of telomerase activity in somatic cell cloned embryos during early development using real-time PCR.the results are as follows:1. Protease concentration is 0.4%,digestion time is 6min,the conditions is better than the other,all 20 oocytes was digested.2.293-cell as the standard and established a standard curve,the correlation coefficient r2=-1,intercept of Y-axis is 45.73,slope is-3.57.3. detection of telomerase activity in somatic cell cloned embryos during early development using real-time PCR.The relativity between CT value and telomerase activity in somatic cell cloned embryos.the Maximum CT value in 8-cell,but telomerase activity is the Minimum;the blastocyst has.the Minimum CT value and the Maximum telomerase activity, Telomerase activity was negatively correlated with the CT value.The CT values was no significant difference in 2-16 cells, but has significant difference in immature oocytes, morula, blastocyst cells.4. compared single cell telomerase activity of oocytes and reconstructed embryo in different stages.the single-cell of immature oocytes has the maximum telomerase activity, the single-cell of blastocyst has the minimum telomerase activity, the single-cell of morula is higher than blastocyst, Telomerase activity from immatured oocytes to the blastocyst was Decreased. |
PDF Full Text Request