Amplificação de gene de lectina a partir de folhas de Bauhinia pentandra (Bong.) D.Dietr.

Abstract

The present study aimed to obtain the amplicon of the lectin gene from leaves of Bauhinia pentandra (Bong.) D.Dietr. Plants of the genus Bauhinia, popularly known as cow’s paw due to the shape of their leaves, are widely used in folk medicine because of their hypoglycemic and diuretic properties. Thus, it is of great interest to investigate the biotechnological potential of their lectins. After collecting young and healthy leaves, the material was immediately stored at –80°C and subsequently subjected to maceration in liquid nitrogen (cell lysis). Nucleic acid extraction was carried out using the CTAB method (cetyltrimethylammonium bromide), followed by RNA removal with RNase A treatment. Primer design and optimization of conventional PCR parameters were then performed. The quality, integrity, and quantity of the DNA obtained were evaluated by agarose gel electrophoresis and spectrophotometry. The resulting amplicon was purified using a specific kit for PCR products. Material with acceptable parameters was obtained through the CTAB method, showing an intact high-molecular-weight band, concentration of 2144 ng/µL, 260/280 ratio = 2.034, and 260/230 ratio = 2.191. After RNase A treatment, the results were 394 ng/µL, 260/280 = 1.807, and 260/230 = 1.361. An amplicon between 750 bp and 1000 bp was obtained, consistent with lectins deposited in sequence databases.