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Construction of a genomic bank of Bradyrhizobium spp. (Rhizobiaceae)

Maria José Valarini^I; Shinjii Tsuyumu^II; João Lúcio de Azevedo^III

^IInstituto de Zootecnia, Caixa Postal 60, 13460-000 Nova Odessa, SP, Brasil. Send correspondence to M.J.V.
^IIFaculty of Agriculture, Shizuoka University, Shizuoka-Shi, Japan
^IIIDepartamento de Genética, ESALQ-USP, Caixa Postal 9, 13400 Piracicaba, SP, Brasil

ABSTRACT

A wild type strain of Bradyrhizobium spp. that associates with Neonotonia wightii (a tropical forage legume) was analysed with molecular biology techniques. A genomic bank of the total DNA from this bacteria was constructed by using the high copy number pUC 119 plasmid, as the cloning vector, established in Escherichia coli HB101 by transformation. The total DNA of Bradyrhizobium was cloned as a collection of Sau 3AI fragments. The hybrid plasmids of the bank contain 10-20kb rhizobia DNA fragments inserted into the lac z region. Assuming that the molecular weight of the Bradyrhizobium genome is about 4,200kb and the average size of the inserts is 15kb, there is more than 95% probability that a nod sequence will be represented in each 10³ transformants, according to the Clarke and Carbon (Cel 9: 91-99, 1976) mathematical expression. Although various chemical and physical methods were used, symbiotic plasmids were not detected in this strain of rhizobia.

Keywords: genomic bank; Bradyrhizobium; Rhizobiaceae.

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