running BLAST (bl2seq) without creating sequence files

Depending on what OS you're running on, you may be able to use something like bash's process substitution. I'm not sure how you'd set that up in Python, but you're basically using a named pipe (or named file descriptor). That won't work if bl2seq tries to seek within the files, but it should work if it just reads them sequentially.

Is this the bl2seq program from BioPerl? If so, it doesn't look like you can do piping to it. You can, however, code your own hack using Bio::Tools::Run::AnalysisFactory::Pise, which is the recommended way of going about it. You'd have to do it in Perl, though.

If this is a different bl2seq, then disregard the message. In any case, you should probably provide some more detail.

How do you know bl2seq does not support piping.? By the way, pipes is an OS feature, not the program. If your bl2seq program outputs something, whether to STDOUT or to a file, you should be able to parse the output. Check the help file of bl2seq for options to output to file as well, eg -o option. Then you can parse the file.

Also, since you are using Python, an alternative you can use is BioPython module.