Method for obtaining nucleic acids from an environment sample, resulting nucleic acids and use in synthesis of novel compounds.

摘要

Preparation of a collection of nucleic acids (I) from organisms in a soil sample, is new. Preparation of a collection of nucleic acids (I) from organisms in a soil sample comprising: (1) milling a dried sample to produce microparticles (MP); (2) suspending these in liquid buffer; (3) extraction of (I) from MP; (4) passing (I)-containing solution through a molecular sieve; (5) passing (I)-enriched fractions through an anion exchange chromatography material; and (6) recovering fractions containing purified (I), is new. Independent claims are also included for the following: (a) processing an environmental sample by: (i) dispersing in liquid; (ii) homogenization of the resulting suspension by gentle mixing; (iii) separation of organisms from other components by density-gradient centrifugation; (iv) lysing separated organisms; (v) extraction of (I); and (vi) purification on a cesium chloride gradient; (b) preparing a collection of recombinant vectors by inserting (I) into a cloning and/or expression vector; (c) new methods for preparing the cloning and expression vectors; (d) a collection of (I) prepared by the new methods; (e) (I) present in the collections of (d); (f) recombinant vectors containing specific (I), prepared by methods (c); (g) collection of recombinant vectors prepared by methods (b) or (c); (h) recombinant vectors present in the collection of (g); (i) recombinant host cells containing (I) of (e) or the vector of (h); (j) collection of cells of (i); (k) detecting a specific (I), or related sequence, in the collection of (j); (l) identifying production of a selected compound (II) in one or more cells of the collection of (j); (m) selected recombinant host cells producing (II) in the collection of (j); (n) producing (II); (o) (II) produced by method (m); (p) polyketide (PK) produced by expressing specified sequences (Ia); (q) determining diversity of (I) in a collection; (r) nucleic acid (Ib) containing a 16S rDNA sequence at least 99% identical with any of 47 specified sequences as given in the specification; (s) producing a type I PK synthase (PKS-I); (t) PKS having any of about 20 specified amino acid sequences as given in the specification; (u) antibodies (Ab) directed against the PKS of (t); (v) detecting PKS-I or its fragments; and (w) kit for method (v).