They all now risk expulsion from the party.
They essentially passed a motion to allow a debate tomorrow on a bill to prevent a 'no deal' Brexit departure from the EU on October 31.
Johnson has indicated he would go to a general election rather than risk a 'no deal' Brexit.
After the result was announced, Johnson says he will table a motion to call for an election if tomorrow's delay on Brexit is passed.
"The leader of the opposition has been begging for an election for two years," he said.
"I don't want an election but if MPs vote tomorrow to compel another pointless delay to Brexit then that would be the only way to resolve this."
The vote defeat comes after Johnson's Conservative government has had their majority of one wiped out during an explosive parliamentary session over the future of Brexit.
The Westminster chamber erupted in a chorus of jeers yesterday as Johnson addressed the packed house in the first session since MPs returned from their summer break.
Speaking 80 years on from the day Britain entered World War II, Johnson said "this country still stands then as now, for democracy, for the rule of law."
"This whole house is united in defending those values."
The comments were met with widespread jeers from fellow MPs, given Johnson has recently asked the Queen to prorogue parliament until October 14 and threatened Conservative MPs with deselection for opposing his Brexit strategy.
Speaker John Bercow was forced to repeatedly call for order during Johnson's short speech.
"Order, order … I ask the house to have some regard for how proceedings are viewed … outside our chamber," he said.
"The Prime Minister is making a statement and that statement should be heard … he will be heard, as will every other member."
Speaking with the sense of a man on a mission, Johnson outlined his recent G7 discussions including a free trade deal for Britain with President Trump in which the NHS would be off the table.
However a major blow came as Conservative MP Phillip Lee crossed the floor to defect to the Liberal Democrats, wiping out the Government's working majority.
Lee said Tuesday he is leaving because "this Conservative Government is aggressively pursuing a damaging Brexit in unprincipled ways."
His decision will weaken Johnson as he tries to prevent parliament from blocking his Brexit plans.
Members of Johnson's own Conservative party are preparing to join opposition politicians in a vote to try to force a delay to Britain's exit from the EU if he cannot secure a divorce deal with Brussels in the next few weeks.
Johnson — who took office less than six weeks ago — has said he wants Britain to leave on October 31 "no ifs or buts" after almost half a century of EU membership.
His decision to suspend Parliament for over a month starting next week so that MPs do not try to hinder his progress has stirred outrage and is being challenged in three separate courts.
On Tuesday, the pound traded at its lowest level since 2016 on fears of the unknown that a "no-deal Brexit" can bring.
