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Background    Eimeria maxima  is one of the most prevalent  Eimeria  species causing avian coccidiosis, and results in huge economic loss to the global poultry industry. Current control strategies, such as anti-coccidial medication and live vaccines have been limited because of their drawbacks. The third generation anticoccidial vaccines including the recombinant vaccines as well as DNA vaccines have been suggested as a promising alternative strategy. To date, only a few protective antigens of  E. maxima  have been reported. Hence, there is an urgent need to identify novel protective antigens of  E. maxima  for the development of neotype anticoccidial vaccines.    Methods  With the aim of identifying novel protective genes of  E. maxima , a cDNA expression library of  E. maxima  sporozoites was constructed using Gateway technology. Subsequently, the cDNA expression library was divided into 15 sub-libraries for cDNA expression library immunization (cDELI) using parasite challenged model in chickens. Protective sub-libraries were selected for the next round of screening until individual protective clones were obtained, which were further sequenced and analyzed.    Results  Adopting the Gateway technology, a high-quality entry library was constructed, containing 9.2 × 10 6  clones with an average inserted fragments length of 1.63 kb. The expression library capacity was 2.32 × 10 7  colony-forming units (cfu) with an average inserted fragments length of 1.64 Kb. The expression library was screened using parasite challenged model in chickens. The screening yielded 6 immune protective genes including four novel protective genes of EmJS-1, EmRP, EmHP-1 and EmHP-2, and two known protective genes of EmSAG and EmCKRS. EmJS-1 is the selR domain-containing protein of  E. maxima  whose function is unknown .  EmHP-1 and EmHP-2 are the hypothetical proteins of  E. maxima.  EmRP and EmSAG are rhomboid-like protein and surface antigen glycoproteins of  E. maxima  respectively, and involved in invasion of the parasite.    Conclusions  Our results provide a cDNA expression library for further screening of T cell stimulating or inhibiting antigens of  E. maxima.  Moreover, our results provide six candidate protective antigens for developing new vaccines against  E. maxima.Electronic supplementary material  The online version of this article (doi:10.1186/s13071-017-2029-4) contains supplementary material, which is available to authorized users.

Identification of immune protective genes of Eimeria maxima through cDNA expression library screening