The Establissshment Of Two-dimensional Electrophoresis And Study On The Antigenicity Of Mycoplasma Ovipneumoniae Y98

Mycoplasma pneumonia of Sheep is one of the most common respriatory disease of sheep and goats. Due to its relatively low death rate, people did not paid much attention to the pathogenic mechanism of this disease. However, recent investigation indicates that the death rate of this disease is increasing, especially among sheep. Today, mycoplasma pneumonia of Sheep is prevailing in a lot of countries and regions around the world and is causing huge economic loss. At the same time, .little is known about its molecular mechanism of pathogen.and the molecular biology development of it is slow. With the test materials of Y98 and PG3, two-way electrophoresis(2-DE)and relevant technology systems study on two mycoplasma proteome is established; and with SDS-PAGE and immunoblott westernblotting, antigen nature of Y98 is explored tentatively. All of the above aim at starting with comstructing the two-way electrophoresis atlas of mycoplasma ovipneumonia proteome study, laying the foundation of molecule biological study which contains ptide quality fingerprint image prepare and acquisition of antigen gene et.al, and genetic engineering vaccine research of sheep pneumonia mycoplasma vaccine, thus hope to control the spread and prevail of this disease fundamentally.The result of SDS-PAGE shows: the whole protein molecular weight range of Y98 is 14kD~100kD, it mainly contains 9 protein strips ,the molecular weight of which are 100 kD, 97kD, 821dD, 74kD, 48kD, 40kD, 35kD, 22 kD, 18kD; the whole protein molecular weight range of HD-1 is also 14kD~100kD, it mainly contains 10 protein strips ,the molecular weight of which are 100kD,97kD, 82kD, 74kD, 48kD, 44kD, 40kD, 35kD, 22kD, 18kD,generally the same as Y98; the whole protein molecular weight range of PG3 is 15kD~123kD, it mainly contains 11 protein strips ,the molecular weight of which are 123kD, 105kD, 73kD, 70kD, 62kD, 38kD, 30.6kD, 29kD, 27.8kD, 23.5kD, 15.6kD,which is obviously different from Y98 and HD-1; the whole protein molecular weight range of FH is 14kD~100kD, it mainly contains 9 protein strips ,the molecular weight of which are 97kD, 79.4kD, 75.9kD, 65kD, 63kD, 53.6kD, 38.2kD, 30kD, 17KD.It is obvious that the bacteria protein strips of HD-1 and Y98 are the same basically, there are only a few different polypeptide .The bacteria protein strips of HD-1 and Y98 have a lot of the same as FH's, but bacteria protein strips of which are greatly different from PG3's .The two-way electrophoresis atlas of proteome shows: the two-way electrophoresis silver dye atlas of Y98 presents 288+9 protein spots , the range of molecular weight is 14 kD-100kD,the range of isoelectric point mainly is 4.436~7.164, which shows the bacteria protein of Y98 are subacidity mainly ; the two-way electrophoresis silver dye atlasof PG3 presents243+ 11 protein spots,the range of molecular weight is 14kD~100kD,the range of isoelectric point mainly is 4.213~7.987,which also proves the bacteria protein of PG3 are subacidity mainly; analyzing the two-way electrophoresis silver dye atlas of Y98 and PG3 by Image Mster 2D Elite 3.01,we find that they have 21 similar protein spots, it may be their common protein .Immunoblott westernblotting of Y98 shows 4 protein strips.the molecular weight of which is 74.2kD, 53.6kD, 40.5kD, 26.5kD,may be the main antigen protein of Y98.Go step futher confirming need further investigation.