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Characterization of DNA Sequences that Confer Complement Resistance in Leishmania chagasi
Author(s) -
DahlinLaborde Rebecca R.,
Scolaro Eric J.,
Romine Nathan M.,
RamerTait Amanda E.,
Lei Soi Meng,
Beetham Jeffrey K.
Publication year - 2008
Publication title -
annals of the new york academy of sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.712
H-Index - 248
eISSN - 1749-6632
pISSN - 0077-8923
DOI - 10.1196/annals.1428.099
Subject(s) - leishmania chagasi , complement (music) , leishmania , chemistry , dna , biology , virology , genetics , leishmaniasis , visceral leishmaniasis , gene , computer science , phenotype , parasite hosting , world wide web , complementation
Serial passage of axenically cultured Leishmania chagasi promastigotes results in a progressive diminution in resistance to complement‐mediated lysis (CML), whereas high CML resistance is seen in infectious metacyclic promastigotes from the sandfly vector as well as metacyclic‐like promastigotes within low‐passage cultures at stationary growth phase. As we previously reported, in a screen seeking to identify novel genes involved in CML resistance: (1) a genomic cosmid library derived from DNA of CML‐resistant L. chagasi promastigotes was transfected into high‐passage (constitutively CML‐sensitive) L. chagasi promastigotes; (2) transformants were screened for acquisition of CML‐resistance; (3) multiple cosmid‐transfectants exhibited partial CML resistance; and (4) the sequence for one of the cosmids (Cosmid 51) was determined. This report extends the analysis of Cosmid 51, and identifies by deletion analysis a subregion of the cosmid insert that is critical to the CML‐resistance phenotype of Cosmid 51 transformants. We also report the sequence determination and initial CML‐resistance activity of another cosmid that also confers partial resistance to CML.