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ãáÎÕ ÇáÈÍË

             Ããßä ÈæÖæÍ ÑæíÉ ÍÒã  ÇáÈÑæÊíä áÈÚÖ ãÓÊÚãÑÇÊ ÇáÚÊÑÇÊ  GL 19, GF 19    ÈÚÏ ÅÓÊÎáÇÕåÇ æÝÕáåÇ ÈÊÞäíÉ ÇáÑÍáÇä ÇáßåÑÈÇÆì áåáÇã ÇáÅßÑíáÃãÇíÏ ÇáãÊÚÏÏ (SDS- PAGE)  æÕÈÛåÇ ÈÇáßæãÇÓí ÇáÃÒÑÞ.  ÃãÇ ÅÎÊÈÇÑ ÊÍáíá ÇááØÎÉ ÇáÛÑÈíÉ  ((Western-blot analysis ÝÞÏ Êã ÅÌÑÇÆå Ýí  ÊÌÑÈÊíä:-

            ÇáÊÌÑÈÉ ÇáÃæáí ßÇäÊ ÈãÞÇÑäÉ ÇáÈÑæÊíä ÇáãæÌæÏ Úáì ÇáÛÔÇÁ ÇáÓÇíÊæÈáÇÒãí áÚÊÑÇÊ ãÌãæÚÉ ÇáãÝØæÑÉ ÇáãÓÊÝØÑÉ( ( M. mycoides clusteræÊãÊ ÈåÐå ÇáÇÎÊÈÇÑÇÊ ãÚÑÝÉ æÊÝÑíÞ ÇáãÝØæÑÉ ÇáÊíÓíÉ ÊÍÊ äæÚ ÅáÊåÇÈ ÑÆÉ ÇáãÇÚÒ  (M.capricolum subsp. Capripneumoniae) Úä   ÈÞíÉ   ÚÊÑÇÊ   ãÌãæÚÉ   ÇáãÝØæÑÉ ÇáãÓÊÝØÑÉ æÐáß áæÌæÏ ÍÒãÉ ÈÑæÊíä ÈÍÌã ÌÒíÆí Èíä 7.6 æ 18 ßíáæ ÏÇáÊæä æáÇ ÊæÌÏ åÐå ÇáÍÒãå Ýí  ÈÞíÉ ÚÊÑÇÊ ÇÝÑÇÏ Êáß ÇáãÌãæÚÉ  ÚáãÇð ÈÃä åÐå ÇáäÊíÌÉ áã íÓÈÞ ÊÓÌíáíåÇ ãä ÞÈá. áÐÇ äæÕí ÈÇÓÊÎÏÇã ÇÎÊÈÇÑ ÇááØÎÉ ÇáÛÑÈíÉ Ýí ÇáÊÚÑÝ Úáì åæíÉ ÇáãÝØæÑÉ ÇáÊíÓíÉ ÊÍÊ äæÚ ÅáÊåÇÈ ÑÆÉ ÇáãÇÚÒ áÊÔÎíÕ ÇáÅáÊåÇÈ ÇáÑÆæí ÇáÈáæÑí ÇáÓÇÑí Ýí ÇáãÇÚÒ. ÝãÓÊÚãÑÇÊ ÇáÚÊÑå  GL 19 ÇáËáÇË áåÐå ÇáãÝØæÑÉ  æÇáãÓÊÎÏãÉ Ýí åÐÇ ÇáÇÎÊÈÇÑ ÃÚØÊ äÊÇÆÌ ãÊØÇÈÞÉ ÍíË ÊÍááÊ ßá ãÓÊÚãÑÉ Åáì ÍæÇáí 35-50 ÍÒã ãä ÇáÈÑæÊíä íÞÚ ãÚÙãåÇ Èíä ÇáÃæÒÇä ÇáÌÒÆíÉ 42.2 æ 132 ßíáæ ÏÇáÊæä. ÃãÇ ÈÞíÉ ÚÊÑÇÊ ÃäæÇÚ ÇáãÌãæÚÉ ÝÞÏ ÃÚØÊ ÃÑÈÚ ÍÒã æÇÖÍÉ ãä  ÇáÈÑæÊíä ÊÑßÒÊ Èíä ÇáÂæÒÇä ÇáÌÒÆíÉ   71 æ 91  ßíáæ ÏÇáÊæä.

ÃãÇ ÇáÊÌÑÈÉ ÇáËÇäíÉ ÝÞÏ ÃÌÑÈÊ áãÞÇÑäÉ ÇáÈÑæÊíä ÇáãÊæÇÌÏ Úáì ÇáÛÔÇÁ ÇáÓáÇíÊæÈáÇÒãí ááãÓÊÚãÑÇÊ ÇáßÈíÑÉ æÇáÕÛíÑÉ ááÚÊÑÊíä GL 19 æGF 19   æßÇä ÇáÝÑÞ ßãíÇð ÍíË ÃÚØÊ ÇáãÓÊÚãÑÇÊ ÇáßÈíÑÉ ãÞÏÇÑÇð ÃßÈÑ ãä ÇáÈÑæÊíä íÞÚ ãÚÙãå Èíä ÇáÃæÒÇä ÇáÌÒÆíÉ 35.1 æ 132 ßíáæ ÏÇáÊæä ÈíäãÇ ÃÚØÊ ÇáãÓÊÚãÑÇÊ ÇáÕÛíÑÉ ãÞÏÇÑÇð ÃÞá ÊÑßÒ Èíä ÇáÃæÒÇä ÇáÌÒÆíÉ 42.2 æ 91 ßíáæ ÏÇáÊæä.

ÇÓÊÎÏãÊ ÊÞäíÉ ÇáÊÞÓíã ÇáãÑÍáì ÈæÇÓØÉ Triton x-114 ßÎØæÉ Çæáí áãÚÑÝÉ áãßæäÇÊ ÇáÓØÍíÉ ááÚÊÑÉ GL 19 ááãÝØæÑÉ ÇáÊíÓíÉ ÊÍÊ äæÚ ÇáÊåÇÈ ÑÆÉ ÇáãÇÚÒ æÇáÊì ÇßÏÊ ÇáÕÝÉ ÇáããíÒÉ ááãÓÊÚãÑÉ ÇáÕÛíÑÉ áåÐå ÇáÚÊÑÉ æÇáÊì æÌÏÊ ÈæÇÓØÉ ÇÎÊÈÇÑ ÊÍáíá ÇááØÎå ÇáÛÑÈíå áßá ÈÑæÊíä ÇáÎáíÉ. ÇÙåÑÊ ßá ÇáÈÑæÊäíÇÊ ÇáÊí ßËÝÊ Ýì ÇáØÈÞÉ ÛíÑ ÇáãÍÈÉ ááãÇÁ ÊÝÇÚáÇð ãäÇÚíÇð Ýí ÇÎÊÈÇÑ ÊÍáíá ÇááØÎÉ ÇáÛÑÈíÉ æåì ÊÞÑíÈÇð äÝÓ ÇáÈÑæÊíäÇÊ ÇáãæáÏÉ ááãäÇÚÉ (æãÓÆæáÉ Úä ÚãáíÉ ÇáÇáÊÕÇÞ) ááãÓÊÚãÑÉ ÇáÕÛíÑÉ ÇáÊí ÊãÊ ÏÑÇÓÇÊåÇ. æåÐå ÇáÍÞíÞÉ ÞÏ ÊÄßÏ ÇáÝÑÖíÉ ÇáÞÇÆáÉ Çä ÇáãÓÊÚãÑÇÊ ÇáÕÛíÑÉ åì ÇáããÑÖÉ.

Summary

Protein bands separated from some colonies of strains GL19 and GF19 were easily visualised on SDS-PAGE, when stained with Coomassie-blue Stain. Western-blot analysis of soluble proteins was done in two experiments:

In the first experiment, the analysis was carried out on members of the Mycoplasma mycoides cluster. A distinct protein band was found between molecular weights 7.6  and 18 KDa, in the three colonies of GL 19 and it was completely absent in all other members of the M. mycoides cluster. Therefore, Western-blot analysis is strongly recommended as an identifying test for M. capricolum subsp. capripneumoniae as well as a differential diagnostic test for CCPP. Moreover, the protein bands of three GL 19 colonies showed 35-50 identical distinct protein bands, the majority of them were between molecular weights 45.2 and 132 KDa, while only 2-4 distinct bands were produced by the other members of the M. mycoides cluster and they were approximately at molecular weights 71-91 KDa. The second experiment compared some of the large and small colonies of strains GL 19 and GF 19. A degree of difference between the two types of colonies was established, confined mainly to quantitative variation of protein bands. The large colonies from both strains provided more deviating patterns and a protein bands of molecular weights of 35.1 to 132 KDa while most of the protein bands from the small colonies were between 45.2 and 91 KDa.

TritonX-114 phase partitioning was employed as first step towards identifying hydrophobic surface constituents of M. capricolum subsp. capripneumoniae strain GL19 which confirmed the characteristic strain features of the small colonies found with whole cell protein in Western-blot analysis. Virtually all of the proteins enriched in the hydrophobic phase exhibited immune reactivity in Western-blot analysis, were actually the same immunogenic proteins (responsible for adherence) of the small colonies of the candidate strain, a fact that might confirm the assumption that the small colonies are the pathogenic types.

 
 
   
 
 

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