You can find details of transformation and conjugation methods in Practical Streptomyces Genetics - see http://www.jic.ac.uk/science/molmicro/Strepmanual/Manual.htm. For knockouts, look up Gust B in Pubmed, especially e.g Gust B, Challis GL, Fowler K, Kieser T, Chater KF. Proc Natl Acad Sci U S A. 2003 Feb 18;100(4):1541-6.
I have been trying genetic modification in A. mediterraneei from last one year via conjugation. I have gone through a number of papers, manual articles and the streptomyces genetic modification book . I did not get success in gene inactivation till date. I am facing some practical problems. After following different conjugation methods published in esteemed journals, most of the time after conjugation I got exconjugants but not in the form of single colonies, it generally grows in form of mat. After re-plating the exconjugants on selective plates, I did not get any positive clone. I am using suicidal, integrative and replicating streptomyces shuttle vectors (pSET152, PKC1218 and pTZ58) for the purpose. It would be a great privilege for me to get your guidance in this aspect.
Hope you might have seen my reply. I had contact a number of scientist in this regard, everbody replied to repeat the experiment again and again. I could not understand where I used to make mistakes, either before conjugation or after conjugation. Here I have limited resources, I do not have any cosmid with A. medi. genome or cells for lamda red recombination. I am greatful to your organization for providing me ET12567 . If you feel resonable,may I discuss this with you . Kindly let me know your view .